HIF-1 alpha Antibody (H1alpha67)

HIF-1 alpha induction by CoCl2 on Caki-1 cell lysate. WB image submitted by a verified customer review.

HIF-1 alpha was detected in immersion fixed DFO treated Hela cells (left) but was not detected in HIF-1 knockout HeLa cells (right) using Mouse Anti-human HIF-1 alpha monoclonal antibody (Catalog # NB100-105) at 25 ug/mL for 3 hours at room temperature. Cells were stained using a NorthernLights (TM) 557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei.

Naive CD4 T cells from WT, VHL-deficient (Vhl KO), or HIF-1 alpha-deficient (HIF-1 alpha KO) mice were differentiated under IL-22-skewing conditions for a total of 60 h. Some cells remained at normoxia for the duration of the culture (N); others were at normoxia for 35 h and then hypoxia (1% O2) for 24 h (H). At 60 h, nuclear extracts were harvested, and HIF-1 alpha and Lamin B1 levels were analyzed by Western blot. Image from verified customer review.

Staining of HIF-1 alpha in human kidney using NB100-105. Renal tubular epithelium showed moderate membranous, cytoplasmic and nuclear staining, and glomeruli showed faint to moderate nuclear staining.

Analysis using the Alexa Fluor (R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in multiple myeloma cells: H929 cells (0.5 x 10^6) were stained with Alexa Fluor 488(R) conjugated HIF-1 alpha antibody (NB100-105AF488). Image courtesy of Dr. Barbara Muz at Washington University in St. Louis School of Medicine.

HIF1alpha/PFKFB3 is upregulated in beta-cells of HIP rats and humans with type 2 diabetes. Representative Western blot of PFKFB3 and HIF1alpha levels in nuclear-enriched- and whole cell extracts from non-diabetic (ND) and T2D donor islets. Data are presented as mean +/- SEM, n = 3 independent biological samples for each group. Statistical significance was analyzed by Student t-test (*p 50 myofibers from 3 mice/group/time point) & stained for Pax7, DAPI, & EdU (A), HIF2A (B), or HIF1A (C). Scale bars: 20 μm. Arrowheads indicate SCs. (D) Number of Pax7+ SCs per myofiber at various time points. (E) Percentage of EdU+ SCs at various time points. (F) Percentage of HIF2A+ SCs at various time points. (G) Percentage of HIF1A+ SCs at various time points. Data represent the mean ± SEM. Image collected & cropped by CiteAb from the following publication (//www.jci.org/articles/view/96208), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - mTORC2 not mTORC1 regulates HIF‐2 alpha , GOT1 in PDAC under prolonged hypoxia. (A & B) HIF‐2 alpha mRNA & protein levels determined by qRT‐PCR (mRNA) & Western blot (protein) after treatment of Panc‐1 & Capan‐2 cells with mTORC1 inhibitor rapamycin for 48 hrs at 3% or 1% O2. beta ‐Actin was used as loading control. (C & D) Panc‐1 & Capan‐2 cells were treated with mTORC1/mTORC2 inhibitor PP242 & cultured for 48 hrs at 20%, 3% or 1% O2, & HIF‐2 alpha , GOT1 mRNA & protein levels were determined by qRT‐PCR (mRNA) & Western blot (protein). beta ‐Actin was used as loading control. Data are presented as mean ± S.D. from three independent experiments. *P 50 myofibers from n = 3 mice) showing that nmGFP+ QSCs were pimonidazole+. Scale bars: 50 μm & 10 μm (insets). Inset images show that pimonidazole signals were relatively enriched in the cytoplasm of QSCs. Arrowheads indicate a QSC; asterisks indicate a myonucleus. (B) Percentage of pimonidazole+ QSCs. (C) Timeline of in vivo CCI-103F labeling in C57BL/6 mice & representative images of uninjured/resting EDL myofibers (n >50 myofibers from 3 mice) showing that nmGFP+ QSCs were CCI-103F+. Arrowheads indicate a QSC; asterisks indicate a myonucleus. Scale bar: 20 μm. (D) Percentage of CCI-103F+ QSCs. (E & F) Representative images of uninjured/resting EDL myofibers from C57BL/6 mice (n >50 myofibers from 6 mice/group) showing that most QSCs were HIF2A+, but HIF1A–. Scale bars: 10 μm. (G) Percentage of HIF1A+ & HIF2A+ QSCs. Data represent the mean ± SEM. Image collected & cropped by CiteAb from the following publication (//www.jci.org/articles/view/96208), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - FDFT1 inhibits glycolysis through suppressing AKT-mTOR-HIF1 alpha pathway in CRC.aFDFT1 overexpression reduced glucose uptake in CT26 cells. bFDFT1 overexpression decreased lactate production via glycolysis in CT26 cells. c ECAR reduced when FDFT1 overexpressed in CT26 cells. d OCR increased when FDFT1 overexpressed in CT26 cells. eFDFT1 knockdown increased glucose uptake in CT26 cells. fFDFT1 knockdown increased lactate production via glycolysis in CT26 cells. g ECAR increased when FDFT1 knocked down in CT26 cells. h OCR decreased when FDFT1 knocked down in CT26 cells. i, mFDFT1 overexpression inhibited protein & mRNA expression of mTOR-targeted glycolytic enzymes, including GLUT1, HK2, PGK1, GPI, & LDHA, in CT26 cells. j, mFDFT1 knockdown increased protein & mRNA expression of mTOR-targeted glycolytic enzymes, including GLUT1, HK2, PGK1, GPI, & LDHA, in CT26 cells. k, nFDFT1 overexpression decreased protein & mRNA expression of AKT, mTOR, & HIF1 alpha . l, nFDFT1 knockdown increased protein & mRNA expression of AKT, mTOR, & HIF1 alpha . o Photograph of dissected tumors (the first line: normal diet, second line: FMD + glucose, third line: FMD, n = 5). p The tumor volumes measured every 2 days after 13th day. The FMD + glucose group can reverse tumor growth inhibition induced by FMD (n = 5; ns: P = 0.1838; P = 0.0001). q The protein level of FDFT1 & mTOR in dissected tumor samples from normal diet group, FMD group & FMD + glucose group measured by western blotting. r The glucose level in these three groups. Error bars, mean ± SD, data are from three independent experiments. Two-sided t tests. *P < 0.05, **P < 0.01, ***P < 0.001, compared w/ control group (or normal diet group). #P < 0.05, ##P < 0.01. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/32313017), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Role of HIF-1 alpha , FOXO3 in hypoxia-induced BNIP3 expression. (A) UCB-hMSCs were incubated with various durations of hypoxia (0–48 h). The protein expressions of HIF-1 alpha & beta -actin were detected by western blot. n = 4. (B) UCB-hMSCs were pretreated with NAC (5 mM) for 30 min prior to hypoxia incubation for 24 h. The protein expressions of HIF-1 alpha , lamin A/C & beta -tubulin in non-nuclear & nuclear fractionized cell samples were assessed by using western blot. n = 3. (C) UCB-hMSCs were immuno-stained with HIF-1 alpha & PI (magnification ×600). Scale bars, 37.5 µm. (D) HIF1A siRNA or NT siRNA was transfected to cells prior to hypoxia treatment for 24 h. The mRNA expression of BNIP3 was analyzed by qPCR. n = 6. (E) The protein expressions of BNIP3 & HIF-1 alpha were detected by western blot. n = 4. (F) NAC (5 mM) was pretreated to UCB-hMSCs prior to hypoxia treatment for 24 h. FOXO3, lamin A/C & beta -tubulin proteins expressions were assessed by western blot. n = 3. (G) FOXO3 siRNA transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. The FOXO3 mRNA expression was measured by qPCR. n = 6. (H) BNIP3, FOXO3 & beta -actin expressions were detected by western blot. n = 3. Western blot data were normalized by beta -actin, & qPCR data were normalized by ACTB mRNA expression level. Lamin A/C & beta -tubulin were used as nuclear & non-nuclear protein controls, respectively. Quantitative data are presented as a mean ± S.E.M. All blots & confocal images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - ROS overproduction mediated apoptosis induced by combined treatment of DDP & HIF-1 alpha in PC-3 xenografts & cell culture. (a) ROS monitored by DHE (red) & nuclei by DAPI (blue) staining in PCa xenografts (scale bars, 50 µm). (b) MDA formation of PCa xenografts was examined after various treatments (c,d). PC-3 cells were treated with DDP, si-HIF-1 alpha plasmid, or both, in the presence or absence of NAC (5 mM) or DHLA (0.25 mM) for 24 h. Total lysates (c) & culture media (d) were used to detect cellular H2O2 level. (e,f) Western analysis for HIF-1 alpha as well as cleaved caspase-3 & PARP in PC-3 cells following various treatments. Data were presented as mean ± SD of three independent experiments. *p < 0.05 versus control group; #p < 0.05 versus si-HIF-1 alpha or DDP group; $p < 0.05 versus DDP/si-HIF-1 alpha group. The original blots are presented in Supplementary Figure 8. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-07973-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - miR-128 downmodulation regulates MAFG increase in hypoxic condition both in vitro & in vivo. C2C12 cells exposed to hypoxia (2% O2) for 4 h, & miR-128 levels (a) determined by RT-qPCR on total RNAs after normalization w/ the small RNU6. Data are expressed as relative to values obtained in untreated control which set equal to 1. Each column in the panels represents the mean ± SD of 3 independent experiments. ∗p < 0.05. Western blotting analysis (b) of HIF-1 alpha , MAFG, & HMOX-1 performed on total extracts from the same cells, as described in Materials & Methods. Tubulin used as a loading control. mRNA levels of HMOX-1 & x-CT genes (c) determined by RT-qPCR on total RNAs, & relative changes calculated by comparing treated cells versus control, after normalization w/ c-ABL. Data reported as relative to values obtained in normoxia, which set equal to 1. Each column in the panel represents the mean ± SD of 3 independent experiments. ∗p < 0.05. Representative laser Doppler analysis (d) of blood flow before & 4 hours after (post 4 h) hindlimb ischemia procedure (n = 3-4 mice/group). Representative Western blot analysis & densitometric analysis (e) of MAFG & HMOX-1 protein levels performed on hindlimb muscle lysates from littermates after sham procedure (sham) or not-ischemic limb (NIL) & ischemic limb (IL) 4 h after femoral artieriectomy. Tubulin protein levels used as loading controls (∗p < 0.05 versus sham; n = 3-4 hindlimb/group). miR-128 levels (f) determined by RT-qPCR on total RNAs after normalization w/ the small RNU6. The values obtained in not-ischemic limb (NIL) set equal to 1. The data are expressed as the mean ± standard error & are representative of 3 independent experiments. ∗p < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29138682), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Effect of ENMD-1198 on HIF-1 alpha & VEGF expression in HCC cells. To determine effects of ENMD-1198 on nuclear HIF-1 alpha expression, HUH-7 cells were incubated for 16 h ± ENMD-1198 & subsequently exposed to hypoxia (1% O2, 4 h). A) Western blot analysis of whole protein showed that ENMD-1198 effectively blunted hypoxic induction of HIF-1 alpha protein. B) In addition, treatment with ENMD-1198 down-regulated constitutive VEGF-A mRNA levels in HCC cells (HepG2), as measured by real-time PCR (* P < 0.01) (n = 3/group). VEGF-A mRNA expression is normalized to beta -actin. C) ELISA analysis for VEGF in culture supernatants (HepG2). Hypoxia (20 h, 1% O2) markedly increased VEGF protein. ENMD-1198 treatment lowered VEGF secretion under hypoxic conditions (*P < 0.05). D) Western blot analysis for VEGF. Cells were incubated under either non-hypoxic, or hypoxic conditions ± ENMD-1198. Treatment with ENMD-1198 slightly lowered the hypoxic induction of VEGF, as determined by densitometry. Bars: SEM. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18651980), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Upregulation of HIF-1 alpha in human PCa. (a) HIF-1 alpha protein was detected by western blot in nonmalignant (RWPE-1 & BPH1) & PCa cell lines (PC-3, DU145, LNCaP, & 22RV1) as indicated. (b–d) Total RNA extracted from RWPE-1, BPH1, PC-3, DU145, LNCaP, & 22RV1 cells was subjected to qRT-PCR for HIF-1 alpha (b), VEGF (c) & GLUT4 (d). (e) The HIF-1 alpha promoter-driven reporter (firefly luciferase) & a control vector (Renilla luciferase) were co-transfected into RWPE-1, BPH1, PC-3, DU145, LNCaP, & 22RV1 cells for measurement of luciferase activity. HIF-1 alpha promoter activity was calculated as a ratio of firefly to Renilla activity. (f) Human normal & malignant tissue (Gleason score 9) sections were probed with HIF-1 alpha antibody (scale bars, 100 µm). (g) Protein expression of HIF-1 alpha , VEGF, & GLUT4 were examined with western blot, in PC-3, DU145, & LNCaP cells after various treatments as indicated. Data are expressed as mean ± SD of seven independent experiments. $p < 0.05 versus RWPE-1 or BPH1 cells or normal tissue. *p < 0.05 versus control group. #p < 0.05 versus si-HIF-1 alpha or DDP group. Original blots are shown in Supplementary Figure 5. C: Ctrl; D: DDP; S: si-HIF-1 alpha ; D/S: DDP/si-HIF-1 alpha . Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-07973-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Cx43-overexpressing cells regulate the expression of HIF-1 alpha , vascular endothelial growth factor (VEGF), & the proliferation of endothelial cells. (a) The expression of Cx43 & HIF-1 alpha was measured in Cx43-overexpressing cells by Western blot analysis. beta -actin expression served as loading controls for & total protein. Inserted values indicated relative proteins expression in comparison with beta -actin; (b) Cx43-overexpressing cells were cotransfected with pCLNCX-6× HRELuc & pTCYLacZ plasmids. At 6 h post-transfection, their luciferase activities were determined & normalized with beta -gal activity. Data shown were the mean ± SD (n = 4); (c) The conditioned medium of Cx43-overexpressing cells was measured by ELISA. Data shown were the mean ± SD (n = 4); & (d) The conditioned medium of Cx43-overexpressing cells reduced the proliferation of endothelial cells. The HEMC-1 cells treated with conditioned medium of Cx43-overexpressing cells were examined the proliferation activity. Cell viability was assessed by the WST-1 assay. Data shown were the mean ± SD (n = 4). * p < 0.05; *** p < 0.001. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/16/1/439), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Prolonged hypoxia increases glutamine metabolism in PDAC cells, & HIF‐2 alpha can promote non‐canonical glutamine metabolism in chronic hypoxic conditions. (A) Time course of glutamine consumption at 1%, 3% or 20% O2, each time data point is an average of triplicate experiments. (B) Panc‐1 & Capan‐2 were incubated for 48 hrs at 1%, 3% or 20% O2, GLS1, GOT1 & GOT2 protein were measured by Western blot. beta ‐Actin was used as loading control. (C) Panc‐1 & Capan‐2 were incubated for 48 hrs at 1%, 3% or 20% O2, GLS1, GOT1 & GOT2 mRNA were measured by qRT‐PCR. Data are presented as mean ± S.D. from three independent experiments. (D & F) Si‐HIF‐2 alpha ‐transfected Panc‐1 & Capan‐2 cultured at 1% or 3%O2 for 48 hrs. The level of HIF‐2 alpha & glutamine metabolism enzymes mRNA & protein were determined by qRT‐PCR (mRNA) & Western blot (protein), & beta ‐actin was used as loading control. Data are presented as mean ± S.D. from three independent experiments. *P < 0.05, **P < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28544376), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF1 alpha & PFKFB3 are protective against hIAPP mediated beta -cell death. a Representative immunoblotting (left) & quantification (right) of the whole cell extract (WCE) from asynchronous INS 832/13 cells treated with either HIF1 alpha inhibitor KC7F2 (10 μM) or PFKFB3 siRNA (75 nM) for 56 h versus control. Cells were transduced with hIAPP adenovirus (75 MOI) or LacZ adenovirus as control (75 MOI) for last 32 h of culture. DMSO or CTRL siRNA, respectively were added to LacZ or hIAPP to equal DMSO or PFKFB3 siRNA used in experimental groups. GAPDH was used as a loading control. Data are presented as mean ± SEM, n = 4, *p < 0.05, **p < 0.01. b Quantification of DNA content distribution after FACS analysis of INS 832/13 cells treated as described in (a). Data are the mean ± SEM, n = 4, *p < 0.05, **p < 0.01. c The frequency of TUNEL positive INS 832/13 cells treated as described in (a). Data are the mean ± SEM, n = 4 in (a) & (b), & n = 3 independent experiments in (c). Statistical significance was analyzed by one-way ANOVA test with Tukey’s post-test (*p < 0.05, **p < 0.01). See Supplementary Fig. 10 for additional supportive data Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31213603), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Immunohistochemical & Western blot analysis of HIF-1 alpha & VEGF expression in OVA-challenged mice.(A & B) Immunostaining for HIF-1 alpha & VEGF in nasal mucosa following the last challenge. Positive HIF-1 alpha signals are brown & are predominantly nuclear & VEGF is predominantly cytoplasmic (magnification 400×, scale bar = 20 µm). (C & D) Representative Western blot analysis showing HIF-1 alpha & VEGF (with beta -actin as a loading control) expression in the nasal mucosa 24 h after the last challenge. All densitometric analyses are presented as the relative ratio of each molecule to beta -actin & the ratio in negative control mice was set to 100. Values represent means±SEM (n = 6 mice). *Significantly different from negative control, p<0.05; #significantly different from positive control, p<0.05. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0048618), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - DC101, evofosfamide, & HIF-1 alpha shRNA treatment of HT1080 sarcomasA. Western blot of HT1080 cells stably transduced with HIF-1 alpha shRNA (sh.HIF-1 alpha ) or scrambled control shRNA (sh.Scr) & grown in 21% oxygen or 1% oxygen. beta -actin serves as loading control. B. Growth of HT1080 cells stably transduced with sh.HIF-1 alpha or sh.Scr in athymic nude mice. Groups were treated with control IgG 20 mg/kg 3 times per week, DC101 20 mg/kg 3 times per week, and/or evofosfamide (Evo) 50 mg/kg 5 times per week. C. Photos of H&E staining, cleaved caspase 3 IF, cleaved caspase 3 & CD31 IF, CD31 IHC, pimonidazole IHC, & HIF-1 alpha IHC. D. Graphs of microvessel density, pimondizale area, & nuclear HIF-1a expression. Bars represent standard deviation. **p < 0.05 compared to all other groups. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.10212), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - WNT11 is induced by hypoxia or hypoxic mimetics in different cell types.(A) Increased Wnt11 mRNA in EMSC adipocytes (Day 12) after hypoxia-mimetic treatments. EMSC adipocytes were treated with CoCl2 (0.1 mM), DFO (0.1 mM) or DMOG (0.1 mM) for 24 hrs. Values were normalized to Tbp mRNA & are expressed relative to control (n = 3). (B,C) Increased Wnt11 mRNA by hypoxia in EMSC preadipocytes & adipocytes (Day 0–12 after differentiation) (B), & C2C12 myoblast & myocyte (Day 0 & 8 after differentiation) (C). Wnt11 mRNA was assessed by quantitative PCR in cells exposed to air (21% O2) or hypoxia (1% O2) for 24 hrs. (n = 4). Values were normalized to Tbp mRNA & are expressed relative to 21% O2 samples (left panel). (D) Immunoblot analyses of HeLa cells under normal air or hypoxia for 24 hrs. (E,F) Induction of Wnt11 by increasing concentrations of DMOG in MDA-MB-231 cells (E) & 4T1 cells (F). (G) EMSCs treated with 0.1 mM DMOG for the indicated times. Wnt11 & Vegf mRNA expression was measured by qPCR & normalized to Tbp mRNA (n = 4). (H) WNT11 protein levels after DMOG treatment normalized to alpha -Tubulin (upper panel; n = 4). Representative immunoblots of EMSCs treated with 0.1 mM DMOG for the indicated times (Lower panel). (I) Protein expression in MDA-MB-231 cells treated with 0.1 mM DMOG. (J) Induction of Wnt11 promoter activity by hypoxia or hypoxia mimetics. pGL3-Wnt11 promoter plasmid was transfected into C2C12 cells. Cells were incubated with DMOG (left panel, n = 4) or under 21% O2 or 1% O2 (right panel, n = 8) for 24 hrs. For panels (A–C,G,H,J), values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels of immunoblotting, laminin, alpha -tubulin, & ERK were used as loading controls, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - CAPE inhibits MCT-induced HIF-1 alpha expression & PDGF-BB production in rats. (A) Rats were treated with PBS or CAPE (5 or 10 mg/kg) from days 14–28 after MCT treatment 14 days (60 mg/kg). Preparation of lung tissues was analyzed by Western blot to determine the levels of HIF-1 alpha protein. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)was used as a loading control. (B) Serum levels of PDGF-BB in rat was assayed by an ELISA kit. (C) Representative images of immunohistochemical staining for HIF-1 alpha in lung sections from the rats. Scale bars, 50 μm. (D) Immunofluorescence staining of alpha-smooth muscle actin ( alpha -SMA) & HIF-1 alpha in lung sections from rats. Scale bars, 50 μm. Data in (A,B) are expressed as mean ± SEM of three independent experiments. *** p < 0.01, as compared with the PBS group. ### p < 0.01, as compared with the rats exposed to MCT alone. Results are representative of three rats per experimental group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30909527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - mTOR expression is inversely correlated with FDFT1 expression.aFDFT1 overexpression inhibited the protein level of mTOR, whereas FDFT1 knockdown increased the protein level of mTOR in CT26 & SW620 cells. bFDFT1 expression & mTOR expression were negatively correlated in CRC patient samples. Scale bar: 200 µm. c The mTOR silencing efficiency of the siRNA in CT26 & SW620 cells was validated by western blotting. d The effect of mTOR silencing on FDFT1 expression level in CT26 was evaluated by western blotting. e The protein level of mTOR when FDFT1 knockdown combined with or without mTOR inhibitor in CT26. f The protein level of pS6k, S6k, pS6, & S6 under the effect of fasting & FDFT1 overexpression in CRC cells. g, h CCK8 proliferation assays showed that the silencing of mTOR decreased the proliferation of CT26 & SW620 cells. k, n The silencing of mTOR reduced glucose uptake & lactate production in CT26 & SW620 cells. i, j, l, o The silencing of mTOR decreased the ECAR & increased the OCR in CT26 & SW620 cells. m The silencing of mTOR decreased the expression of AKT, HIF1 alpha , & proteins encoded by relevant glycolytic genes, such as GLUT1, HK2, LDHA, GPI, PGK1, in CT26 & SW620 cells. p–t Based on TCGA data set analysis, mTOR expression was positively correlated with AKT1, HIF1 alpha , GLUT1, HK2, & LDHA expression. Error bars, mean ± SD, the data are from three independent experiments. Two-sided t tests. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the control group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32313017), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Chromatin Immunoprecipitation: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF-1 alpha -mediated repression of ENT1 expression in Caki-1 cells under hypoxia. Comparison of HIF-1 alpha & HIF-2 alpha protein expression in human matched renal normal-tumor samples (n=11) (A) & 3 RCC cell lines (786-O, 769-P, Caki-1) under normoxia or hypoxia for 6 h (B). (C) Schematic diagram of two hypoxia response elements (HRE) in the promoter of ENT1. (D) CHIP-qPCR analysis of HIF-1 alpha at the promoter of ENT1 in Caki-1 cells, exposed under normoxia or hypoxia for 48 h. Two different HIF-1 alpha antibodies (610958, BD Biosciences; NB100-105, Novus) were used to pull down HIF-1 alpha . The protein expression level of HIF-1 alpha (E) & mRNA expression level of SLC29A1 (F) in Caki-1 under normoxia or hypoxia & HIF-1 alpha knockout-Caki-1 cells. (G) The expression level of ARNT1 (HIF-1 beta ) was detected in Caki-1, transfected with ARNT1 specific siRNAs or NC. Student's t test (two-tailed) was used. (H, I) The expression level of ENT1 in Caki-1, transfected with ARNT1 specific siRNAs or NC, under normoxia or hypoxia. Oneway-ANOVA analysis was used. The results are expressed as mean ± SEM (n=3). ***P<0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32206108), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Upregulation of HIF-1 alpha in human PCa. (a) HIF-1 alpha protein was detected by western blot in nonmalignant (RWPE-1 & BPH1) & PCa cell lines (PC-3, DU145, LNCaP, & 22RV1) as indicated. (b–d) Total RNA extracted from RWPE-1, BPH1, PC-3, DU145, LNCaP, & 22RV1 cells was subjected to qRT-PCR for HIF-1 alpha (b), VEGF (c) & GLUT4 (d). (e) The HIF-1 alpha promoter-driven reporter (firefly luciferase) & a control vector (Renilla luciferase) were co-transfected into RWPE-1, BPH1, PC-3, DU145, LNCaP, & 22RV1 cells for measurement of luciferase activity. HIF-1 alpha promoter activity was calculated as a ratio of firefly to Renilla activity. (f) Human normal & malignant tissue (Gleason score 9) sections were probed with HIF-1 alpha antibody (scale bars, 100 µm). (g) Protein expression of HIF-1 alpha , VEGF, & GLUT4 were examined with western blot, in PC-3, DU145, & LNCaP cells after various treatments as indicated. Data are expressed as mean ± SD of seven independent experiments. $p < 0.05 versus RWPE-1 or BPH1 cells or normal tissue. *p < 0.05 versus control group. #p < 0.05 versus si-HIF-1 alpha or DDP group. Original blots are shown in Supplementary Figure 5. C: Ctrl; D: DDP; S: si-HIF-1 alpha ; D/S: DDP/si-HIF-1 alpha . Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-07973-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Upregulation of HIF-1 alpha in the spleen restricts CD8+ T cell expansion.(A-B) Mice were infected with 2x107 amastigotes intravenously & euthanized at various time points after infection. HIF-1 alpha expression in splenocytes was assessed by real-time PCR (A) & immunoblot analysis (B). (C-F) 2x104 OT-I CD8+ T cells were adoptively transferred into recipient mice a day prior to infection with ovalbumin-transgenic (PINK) L. donovani amastigotes. (C) Graphs represent the average number of OT-I CD8+ T cells found in the spleen from Hif1aflox/flox & Hif1a+/-mice at d7 & d14 p.i.. (D) Percentage of OT-I CD8+ T cells at d7 & d14 p.i.. (E) Percentage of gated OT-I CD8+ expressing CD62Llo/int & negative for CD127. (F) Percentage of OT-I CD8+ T cells that were CD44+ CD127+CD62Lhi. All data represent mean ± SEM of one of 3 independent experiments, n = 5. * denotes p<0.05 & ** denotes p<0.01. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1004938), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Ccl7 is a HIF1 alpha direct target gene.a qPCR analysis of Ccl7 mRNA expression in Hif1afl/fl & Hif1a delta SMC VSMCs treated with vehicle, CoCl2, normoxia, or hypoxia (2% O2) for 6, 12, & 24 h. *P < 0.05, **P < 0.01, ***P < 0.001, n = 6 (independent experiments) per group. VSMCs isolated from Hif1afl/fl mice & Hif1a delta SMC mice were infected with oxygen-stable HIF1 alpha -expressing lentivirus, & then treated with Ang II for 24 h, bCcl7 mRNA was measured by qPCR & c CCL7 protein was detected by ELISA. d qPCR analysis of Ccl7 mRNA expression in vehicle or Ang II-treated Hif2afl/fl & Hif2a delta SMC VSMCs. e Schematic diagram of the mouse Ccl7 promoter illustrating the HREs in the regulatory region; the upstream regions were numbered in relation to the transcription initiation site. f Luciferase-reporter constructs under the control of the mouse Ccl7 promoter. HEK293T human embryonic kidney cells transiently transfected with the luciferase construct, & cotransfected with empty vector or HIF1a expression plasmids. Standard dual-luciferase assays were performed. EV, empty vector. **P < 0.01, n = 3. g, h ChIP assays of vehicle or Ang II-treated wild-type VSMCs using HIF1 alpha or HIF2 alpha antibodies. Data were normalized to input. *P < 0.05, **P < 0.01, n = 6 per group. i, j ChIP assays of vehicle or Ang II-treated Hif1afl/fl & Hif1a delta SMC VSMCs using HIF1 alpha antibody. Data were normalized to input. *P < 0.05, **P < 0.01, n = 6 per group. Statistical significance was determined by one-way ANOVA test followed by the unpaired t-test Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31320613), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - WNT11 is induced by hypoxia or hypoxic mimetics in different cell types.(A) Increased Wnt11 mRNA in EMSC adipocytes (Day 12) after hypoxia-mimetic treatments. EMSC adipocytes were treated with CoCl2 (0.1 mM), DFO (0.1 mM) or DMOG (0.1 mM) for 24 hrs. Values were normalized to Tbp mRNA & are expressed relative to control (n = 3). (B,C) Increased Wnt11 mRNA by hypoxia in EMSC preadipocytes & adipocytes (Day 0–12 after differentiation) (B), & C2C12 myoblast & myocyte (Day 0 & 8 after differentiation) (C). Wnt11 mRNA was assessed by quantitative PCR in cells exposed to air (21% O2) or hypoxia (1% O2) for 24 hrs. (n = 4). Values were normalized to Tbp mRNA & are expressed relative to 21% O2 samples (left panel). (D) Immunoblot analyses of HeLa cells under normal air or hypoxia for 24 hrs. (E,F) Induction of Wnt11 by increasing concentrations of DMOG in MDA-MB-231 cells (E) & 4T1 cells (F). (G) EMSCs treated with 0.1 mM DMOG for the indicated times. Wnt11 & Vegf mRNA expression was measured by qPCR & normalized to Tbp mRNA (n = 4). (H) WNT11 protein levels after DMOG treatment normalized to alpha -Tubulin (upper panel; n = 4). Representative immunoblots of EMSCs treated with 0.1 mM DMOG for the indicated times (Lower panel). (I) Protein expression in MDA-MB-231 cells treated with 0.1 mM DMOG. (J) Induction of Wnt11 promoter activity by hypoxia or hypoxia mimetics. pGL3-Wnt11 promoter plasmid was transfected into C2C12 cells. Cells were incubated with DMOG (left panel, n = 4) or under 21% O2 or 1% O2 (right panel, n = 8) for 24 hrs. For panels (A–C,G,H,J), values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels of immunoblotting, laminin, alpha -tubulin, & ERK were used as loading controls, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - CAPE inhibits MCT-induced HIF-1 alpha expression & PDGF-BB production in rats. (A) Rats were treated with PBS or CAPE (5 or 10 mg/kg) from days 14–28 after MCT treatment 14 days (60 mg/kg). Preparation of lung tissues was analyzed by Western blot to determine the levels of HIF-1 alpha protein. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)was used as a loading control. (B) Serum levels of PDGF-BB in rat was assayed by an ELISA kit. (C) Representative images of immunohistochemical staining for HIF-1 alpha in lung sections from the rats. Scale bars, 50 μm. (D) Immunofluorescence staining of alpha-smooth muscle actin ( alpha -SMA) & HIF-1 alpha in lung sections from rats. Scale bars, 50 μm. Data in (A,B) are expressed as mean ± SEM of three independent experiments. *** p < 0.01, as compared with the PBS group. ### p < 0.01, as compared with the rats exposed to MCT alone. Results are representative of three rats per experimental group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30909527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Role of HIF-1 alpha , FOXO3 in hypoxia-induced BNIP3 expression. (A) UCB-hMSCs were incubated with various durations of hypoxia (0–48 h). The protein expressions of HIF-1 alpha & beta -actin were detected by western blot. n = 4. (B) UCB-hMSCs were pretreated with NAC (5 mM) for 30 min prior to hypoxia incubation for 24 h. The protein expressions of HIF-1 alpha , lamin A/C & beta -tubulin in non-nuclear & nuclear fractionized cell samples were assessed by using western blot. n = 3. (C) UCB-hMSCs were immuno-stained with HIF-1 alpha & PI (magnification ×600). Scale bars, 37.5 µm. (D) HIF1A siRNA or NT siRNA was transfected to cells prior to hypoxia treatment for 24 h. The mRNA expression of BNIP3 was analyzed by qPCR. n = 6. (E) The protein expressions of BNIP3 & HIF-1 alpha were detected by western blot. n = 4. (F) NAC (5 mM) was pretreated to UCB-hMSCs prior to hypoxia treatment for 24 h. FOXO3, lamin A/C & beta -tubulin proteins expressions were assessed by western blot. n = 3. (G) FOXO3 siRNA transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. The FOXO3 mRNA expression was measured by qPCR. n = 6. (H) BNIP3, FOXO3 & beta -actin expressions were detected by western blot. n = 3. Western blot data were normalized by beta -actin, & qPCR data were normalized by ACTB mRNA expression level. Lamin A/C & beta -tubulin were used as nuclear & non-nuclear protein controls, respectively. Quantitative data are presented as a mean ± S.E.M. All blots & confocal images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Cx43-knockdown cells regulate the expression of HIF-1 alpha , c-Src, VEGF & the proliferation of endothelial cells. (a) The expression of Cx43, c-Src, & HIF-1 alpha was measured in Cx43-knocknown cells by Western blot analysis. beta -actin expression served as loading controls for & total protein. Inserted values indicated relative protein expression in comparison with beta -actin; (b) Cx43-knockdown cells were cotransfected with pCLNCX-6× HRELuc & pTCYLacZ plasmids. Then, cells were treated with 100 ng/μL PP2 or DMSO for 1 h. At 6 h post-transfection, their luciferase activities were determined & normalized with beta -gal activity. Data shown were the mean ± SD (n = 4); (c) The conditioned medium of Cx43-knockdown cells was measured by ELISA. Data shown were the mean ± SD (n = 4); (d) The conditioned medium of Cx43-knockdown cells reduced the proliferation of endothelial cells. The HEMC-1 cells treated with conditioned medium of Cx43-knockdown cells were examined for proliferation activity. Cell viability was assessed by the WST-1 assay. Data shown were the mean ± SD (n = 4). * p < 0.05; ** p < 0.01; *** p < 0.001. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/16/1/439), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Involvement of CBP in BNIP3 expression regulated by HIF-1 alpha & FOXO3 under hypoxia. (A) UCB-hMSCs were incubated with hypoxia condition for 24 h. Co-immunoprecipitation of HIF-1 alpha & FOXO3 with IgG & CBP were shown in left panel. IgG was used as a negative control. The total protein expressions of HIF-1 alpha , FOXO3, CBP & beta -actin in lysate were shown in right panel. n = 3. (B) CBP (20 μM) was pretreated to UCB-hMSCs, & cells were incubated with hypoxia for 24 h. The BNIP3 mRNA expression level was analyzed by qPCR. n = 6. (C) BNIP3 & beta -actin protein expressions were analyzed by western blot. Data represent mean ± S.E. n = 4. (D, E) Sample DNA was immuno-precipitated with RNA polymerase, IgG, HIF-1 alpha & FOXO3 specific antibodies. CHIP (top panel) & lysate (bottom panel) samples were amplified with the primers of GAPDH & BNIP3 promoters. Quantitative CHIP data was analyzed by qPCR, & shown in the right panel. n = 4. Western blot data were normalized by beta -actin, & qPCR data were normalized by ACTB mRNA expression level. Quantitative data are presented as a mean ± S.E.M. All blot images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Precision quantitative image analysis workflow for the characterization of functional ITH.(a) Representative areas with positive immunohistochemical (IHC) staining for HIF-1 alpha , HIF-2 alpha , phospho-mTOR S2448, phospho-S6RP S235/236 & Ki-67. Only nuclear HIF-1 alpha & HIF-2 alpha expression was included in the analysis. Scale bar, 100 μm. (b) Overview of the quantitative IHC analysis process using a representative image of a whole-slide scanned pT1M1 tumour after phospho-mTOR S2448 staining. The image was overlaid with a virtual grid consisting of 1 mm2 squares that defines the tumour regions to be analysed. Non-cancerous tissue was subtracted manually (dark grey) & a positive pixel count was performed on the corrected area. Positivity per square was expressed as percentage of positive pixels. If a square contained non-tumorous tissue, the positivity was normalized to the corrected square area. Computed values were depicted as a heat map. Coloured squares represent tumour, whereas open squares indicate completely non-tumorous areas. Squares defined as peripheral tumour zone are marked by a black line. Scale bar, 1 cm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27291893), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - ROS overproduction mediated apoptosis induced by combined treatment of DDP & HIF-1 alpha in PC-3 xenografts & cell culture. (a) ROS monitored by DHE (red) & nuclei by DAPI (blue) staining in PCa xenografts (scale bars, 50 µm). (b) MDA formation of PCa xenografts was examined after various treatments (c,d). PC-3 cells were treated with DDP, si-HIF-1 alpha plasmid, or both, in the presence or absence of NAC (5 mM) or DHLA (0.25 mM) for 24 h. Total lysates (c) & culture media (d) were used to detect cellular H2O2 level. (e,f) Western analysis for HIF-1 alpha as well as cleaved caspase-3 & PARP in PC-3 cells following various treatments. Data were presented as mean ± SD of three independent experiments. *p < 0.05 versus control group; #p < 0.05 versus si-HIF-1 alpha or DDP group; $p < 0.05 versus DDP/si-HIF-1 alpha group. The original blots are presented in Supplementary Figure 8. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-07973-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Regulation of OA development by HIF-1 alpha . Plain radiographs of the entire bodies at 8 weeks of age (A), & knee joints 8 weeks after OA surgery (B) of Cntl & cKO littermates. Scale bars, 10 mm & 1 mm, respectively. (C) Safranin O & TUNEL staining’s of Cntl & KO knee joints at 8 weeks after OA surgery (n = 9 & 11, respectively). Tamoxifen induction was performed at 7 weeks. Inset boxes in top images indicate the regions of high magnification Safranin O & TUNEL images. Inset boxes in middle images indicate the regions of immunofluorescence in (G). White dotted lines in TUNEL images indicate the cartilage area. Scale bars, 400 µm & 100 µm, respectively. (D) OARSI (Osteoarthritis Research Society International) score of OA development. Lines show the mean ± SD. *P < 0.05. (E) Rate of TUNEL-positive cells in the cartilage area. Bars show the mean ± SD. *P < 0.05. (F) mRNA levels of Hif1a in articular cartilage tissues of Cntl & KO. Bars show the mean ± SD. *P < 0.05. (G) Immunofluorescence of Hif1a, Mmp13, Hif2a, I kappa B alpha , Ser 32 & 36 dual phosphorylated I kappa B alpha , Rela, & C1qtnf3. Scale bars, 20 µm. The percentage of positive cells in the immunofluorescence is shown below. *P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32214220), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - A beta facilitates HIF1 alpha synthesis & autophagy inhibition via mTOR activation. (A) SK-N-MC cells exposed to A beta (5 μM) for 0–48 h. HIF1 alpha & beta -actin expression analyzed by WB. n = 3. (B) Cells pretreated w/ NAC (1 mM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expression analyzed by WB. n = 3. (C,E) Cells incubated w/ rapamycin (10 nM) for 30 min prior to A beta treatment for 24 h. Phosphorylation of 4EBP1 (Thr 37/46) & 4EBP1, phosphorylation of p70S6K1 (Thr 389), HIF1 alpha & beta -actin analyzed by WB. n = 6. (D) Protein samples immunoprecipitated by eukaryotic translation initiation factor 4E (eIF4E) antibody-conjugated protein A/G agarose beads. Samples blotted w/ 4EBP1 & eIF4E-specific antibodies. n = 3. (F) Cells exposed to PF4708671 (10 μM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expression detected by WB. n = 6. (G) Cells exposed to cycloheximide (4 μM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expressions detected by WB. n = 6. (H) Cells pretreated w/ rapamycin (10 nM) for 30 min, incubated w/ A beta for 24 h & analyzed by WB w/ LC3, p62 & beta -actin specific antibodies. n = 3–6. (I) LC3 puncta visualized by confocal microscopy. Presented results merged images. Green & red fluorescents indicate LC3 & PI respectively. Scale bars, 50 μm (magnification × 600). (J) Cells pretreated w/ trehalose (10 μM) for 30 min prior to A beta treatment for 24 h. Cytotoxicity measured by MTT assay at an absorbance of 545 nm using a microplate reader. Data present the mean ± SE. n = 6. (K) Cell viability measured by trypan blue exclusion assay. Data presented as a mean ± SE. n = 6. Each blot image presented as representative image. *p < 0.05 vs. control, #p < 0.05 vs. A beta treatment. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28790888), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - TSA reduces HIF-1 alpha & VEGF expression & up-regulates the expression of PEDF.Real-time PCR (A) & Western blot assays (B) were performed on RPE cells treated with 0–0.5 μM TSA & 150 μM CoCl2. (A) Cells were treated with 0–0.5 μM TSA for 14 h & then co-treated with 150 μM CoCl2 for 6 h for the analysis of gene expression by real-time PCR. Changes in HIF-1 alpha mRNA levels were not statistically significant. CoCl2 causes a fourfold enhancement of VEGF mRNA expression; but at 0.5 μM TSA, the mRNA level of VEGF reduces to less than half of that in cells treated with CoCl2 only. TSA induces a statistically significant increase in the mRNA level of PEDF. (B) Cells were treated with 0–0.5 μM TSA for 18 h & then co-treated with 150 μM CoCl2 for 6 h for Western blot analysis. TSA reduces the CoCl2-induced HIF-1 alpha & VEGF protein levels by 4.3-fold & 5.7-fold, respectively, & up-regulates PEDF protein level by threefold. (C) Densitometry data for Western blot of HIF-1 alpha , VEGF & PEDF. (*: t test p<0.05; **: t test p<0.01). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0120587), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - A beta facilitates HIF1 alpha synthesis & autophagy inhibition via mTOR activation. (A) SK-N-MC cells exposed to A beta (5 μM) for 0–48 h. HIF1 alpha & beta -actin expression analyzed by WB. n = 3. (B) Cells pretreated w/ NAC (1 mM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expression analyzed by WB. n = 3. (C,E) Cells incubated w/ rapamycin (10 nM) for 30 min prior to A beta treatment for 24 h. Phosphorylation of 4EBP1 (Thr 37/46) & 4EBP1, phosphorylation of p70S6K1 (Thr 389), HIF1 alpha & beta -actin analyzed by WB. n = 6. (D) Protein samples immunoprecipitated by eukaryotic translation initiation factor 4E (eIF4E) antibody-conjugated protein A/G agarose beads. Samples blotted w/ 4EBP1 & eIF4E-specific antibodies. n = 3. (F) Cells exposed to PF4708671 (10 μM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expression detected by WB. n = 6. (G) Cells exposed to cycloheximide (4 μM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expressions detected by WB. n = 6. (H) Cells pretreated w/ rapamycin (10 nM) for 30 min, incubated w/ A beta for 24 h & analyzed by WB w/ LC3, p62 & beta -actin specific antibodies. n = 3–6. (I) LC3 puncta visualized by confocal microscopy. Presented results merged images. Green & red fluorescents indicate LC3 & PI respectively. Scale bars, 50 μm (magnification × 600). (J) Cells pretreated w/ trehalose (10 μM) for 30 min prior to A beta treatment for 24 h. Cytotoxicity measured by MTT assay at an absorbance of 545 nm using a microplate reader. Data present the mean ± SE. n = 6. (K) Cell viability measured by trypan blue exclusion assay. Data presented as a mean ± SE. n = 6. Each blot image presented as representative image. *p < 0.05 vs. control, #p < 0.05 vs. A beta treatment. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28790888), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Role of HIF-1 alpha , FOXO3 in hypoxia-induced BNIP3 expression. (A) UCB-hMSCs were incubated with various durations of hypoxia (0–48 h). The protein expressions of HIF-1 alpha & beta -actin were detected by western blot. n = 4. (B) UCB-hMSCs were pretreated with NAC (5 mM) for 30 min prior to hypoxia incubation for 24 h. The protein expressions of HIF-1 alpha , lamin A/C & beta -tubulin in non-nuclear & nuclear fractionized cell samples were assessed by using western blot. n = 3. (C) UCB-hMSCs were immuno-stained with HIF-1 alpha & PI (magnification ×600). Scale bars, 37.5 µm. (D) HIF1A siRNA or NT siRNA was transfected to cells prior to hypoxia treatment for 24 h. The mRNA expression of BNIP3 was analyzed by qPCR. n = 6. (E) The protein expressions of BNIP3 & HIF-1 alpha were detected by western blot. n = 4. (F) NAC (5 mM) was pretreated to UCB-hMSCs prior to hypoxia treatment for 24 h. FOXO3, lamin A/C & beta -tubulin proteins expressions were assessed by western blot. n = 3. (G) FOXO3 siRNA transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. The FOXO3 mRNA expression was measured by qPCR. n = 6. (H) BNIP3, FOXO3 & beta -actin expressions were detected by western blot. n = 3. Western blot data were normalized by beta -actin, & qPCR data were normalized by ACTB mRNA expression level. Lamin A/C & beta -tubulin were used as nuclear & non-nuclear protein controls, respectively. Quantitative data are presented as a mean ± S.E.M. All blots & confocal images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF1 alpha protein is suppressed by ED71 but not by 1,25(OH)2D3.(A) Western analysis of Raw264.7 cells cultured in hypoxic conditions with or without 10−7 M of ED71 or 1,25(OH)2D3 (1,25D). (B) Hif1 alpha mRNA levels in Raw264.7 cells cultured in hypoxic conditions were analyzed by realtime PCR in the presence or absence of 10−7 M ED71 or 1,25(OH)2D3. Data represent mean Hif1 alpha expression relative to that of Actb ± SD (n = 5). (C) Levels of VDR transcripts in Raw264.7 cells transfected with shRNA targeting the VDR (shVDR) or control shRNA (Control) were determined by realtime PCR. Data represent mean VDR expression relative to that of Actb ± SD (n = 5). (D) Western analysis of control (shControl) or VDR-suppressed (shVDR#1 or shVDR#2) Raw264.7 transformants cultured in hypoxic conditions with ED71 or 1,25(OH)2D3 (1,25D), both at 10−7 M. (E) M-CSF-dependent Ctsk Cre/Hifflox/flox cells were cultured in normoxic conditions to suppress HIF1 alpha in the presence of M-CSF (50 ng/ml) plus RANKL (25 ng/ml) with either ED71 or 1,25(OH)2D3 (1,25D) both at 10−7M for 4 days. Expression of Ctsk & NFATc1 was then assessed by realtime PCR. Data represent mean Ctsk or NFATc1 expression relative to that of Actb ± SD (n = 5). *P<0.05; **P<0.01; ***P<0.001. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0111845), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Lack of HIF-P4H-1 increases the amount of p53. (a) Western blot analysis of p53 in wt & Hif-p4h-1−/− MEFs cultured in 21% or 1% O2 for 24 h. (b,c) Western blot analysis of p53 in Hif-p4h-1 & scrambled (Scrm) siRNA transfected HEK293 cells (b) & wt MEFs treated with 50 µM FG4497 for 24 h (c). (d) qPCR analysis of p53 mRNA in wt & Hif-p4h-1−/− MEFs cultured in 21% or 1% O2 for 24 h. (e) Analysis of p53 protein turnover rate. Hif-p4h-1−/− & wt MEFs were treated with 200 μg/ml of cycloheximide for the indicated time points & whole cell lysates were blotted for p53. (f) Western blot analysis of MDM2 in wt & Hif-p4h-1−/− MEFs cultured in 21% or 1% O2 for 24 h. (g) Western blot analysis of p53 & MDM2 in wt MEFs treated with 10 μM nutlin-3a for 24 h with or without overexpression of human HIF-P4H-1 (OE). (h,i) Western blot analysis of HIF1 alpha in wt & Hif-p4h-1−/− MEFs cultured in 21% or 1% O2 for 24 h (h) & in scrambled & Hif-p4h-1 siRNA transfected HEK293 cells (i). (j) Western blot analysis of ubiquitination of p53 in Hif-p4h-1 & scrambled siRNA transfected HEK293 cells. The cells were transfected with HA-ubiquitin & endogenous p53 was immunoprecipitated followed by Western blotting with anti-HA & anti-P53 antibodies. (k) Western blot analysis of p53 in Hif-p4h-1−/− & wt MEFs were treated with or without 10 μM MG132 for 5 h. Data are presented as representative Western blots & as mean ± s.d., n = at least 3 individual MEF isolates or experiments. *P < 0.05, **P < 0.01 & ***P < 0.001, two-tailed Student’s t-test. Unprocessed original scans of blots are shown in Supplementary Fig. 5. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-17376-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - NDRG1 was regulated by HIF‐1/2 alpha . (A‐E) The protein expression of the shown genes was detected by Western blot, respectively; beta ‐actin was used as the loading control. (A) RCC4 & 786‐O cells (B) were infected with VHL vector (VHL) or empty vector (EV). (C) Caki‐1 cells were transfected with VHL shRNAs (shVHL) or non‐specific control (NC). (D) RCC4 cells were infected with HIF‐1 alpha siRNAs (siHIF‐1 alpha ) or non‐specific control (NC). (E) 786‐O cells were transfected with HIF‐2 alpha shRNAs (shHIF‐2 alpha ) or non‐specific control (NC). (F) Representative images of HIF‐1 alpha , HIF‐2 alpha & NDRG1 protein expression from the IHC staining of human clear cell renal cell cancer specimens. Patient I: The protein levels of HIF‐1 alpha , HIF‐2 alpha & NDRG1 were low. Patient II: The protein levels of HIF‐1 alpha , HIF‐2 alpha & NDRG1 were medium. Patient III: The protein levels of HIF‐1 alpha , HIF‐2 alpha & NDRG1 were high (scale bar, 50 μm). (G & H) The protein level of NDRG1 is significantly correlated with HIF‐1 alpha (G) & HIF‐2 alpha (H). The protein levels were evaluated according to IHC scores, representing very low (score 0‐2), low (score 3‐4), high (score 5‐8) & strong (score 9‐12). The subjects were divided into four groups according to the IHC scores of HIFs in the tumours. (I) The protein level of NDRG1 is significantly correlated with the average of HIF‐1 alpha & HIF‐2 alpha in ccRCC tissues. The protein levels are evaluated according to IHC scores, representing very low (score < 3), low (score < 5), high (score < 8) & strong (score 8‐12). The subjects were divided into four groups according to the IHC scores of the average of HIF‐1 alpha & HIF‐2 alpha . Significance p‐values & r values according to Spearman's rank correlation Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32537867), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - WNT11 is induced by hypoxia or hypoxic mimetics in different cell types.(A) Increased Wnt11 mRNA in EMSC adipocytes (Day 12) after hypoxia-mimetic treatments. EMSC adipocytes were treated with CoCl2 (0.1 mM), DFO (0.1 mM) or DMOG (0.1 mM) for 24 hrs. Values were normalized to Tbp mRNA & are expressed relative to control (n = 3). (B,C) Increased Wnt11 mRNA by hypoxia in EMSC preadipocytes & adipocytes (Day 0–12 after differentiation) (B), & C2C12 myoblast & myocyte (Day 0 & 8 after differentiation) (C). Wnt11 mRNA was assessed by quantitative PCR in cells exposed to air (21% O2) or hypoxia (1% O2) for 24 hrs. (n = 4). Values were normalized to Tbp mRNA & are expressed relative to 21% O2 samples (left panel). (D) Immunoblot analyses of HeLa cells under normal air or hypoxia for 24 hrs. (E,F) Induction of Wnt11 by increasing concentrations of DMOG in MDA-MB-231 cells (E) & 4T1 cells (F). (G) EMSCs treated with 0.1 mM DMOG for the indicated times. Wnt11 & Vegf mRNA expression was measured by qPCR & normalized to Tbp mRNA (n = 4). (H) WNT11 protein levels after DMOG treatment normalized to alpha -Tubulin (upper panel; n = 4). Representative immunoblots of EMSCs treated with 0.1 mM DMOG for the indicated times (Lower panel). (I) Protein expression in MDA-MB-231 cells treated with 0.1 mM DMOG. (J) Induction of Wnt11 promoter activity by hypoxia or hypoxia mimetics. pGL3-Wnt11 promoter plasmid was transfected into C2C12 cells. Cells were incubated with DMOG (left panel, n = 4) or under 21% O2 or 1% O2 (right panel, n = 8) for 24 hrs. For panels (A–C,G,H,J), values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels of immunoblotting, laminin, alpha -tubulin, & ERK were used as loading controls, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Fe, Ma, Pm, Pm, Rb, Sh, Xp
• Applications: WB, Simple Western, ChIP, ELISA, Flow, Func, GS, ICC/IF, IHC, IP, NULL, WB, ChIP, IHC, KD, KO
• Clone: H1alpha67
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

HIF-1 alpha Antibody (H1alpha67) Summary

• Immunogen: This HIF-1 alpha Antibody (H1alpha67) was developed against a fusion protein containing amino acids 432 - 528 of human HIF-1 alpha [Uniprot# Q16665].
• Localization: HIF-1 is a nuclear protein
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: HIF1A
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 1-5 ug/IP
  • Chromatin Immunoprecipitation 1 - 5 ug/IP. Use reported in scientific literature
  • ELISA 1:100 - 1:2000. Use reported in scientific literature (PMID 20042684)
  • Flow Cytometry 1:10 - 1:1000
  • Gel Super Shift Assays 1:1 - 1:100. Use reported in scientific literature (PMID 22411794)
  • Immunoassay reported in scientific literature (PMID 26147748)
  • Immunoblotting reported in multiple pieces of scientific literature
  • Immunocytochemistry/ Immunofluorescence 1:50
  • Immunohistochemistry Free-Floating reported in scientific literature (PMID 33242463)
  • Immunohistochemistry 1:20-1:50
  • Immunohistochemistry-Frozen 1:20-1:50
  • Immunohistochemistry-Paraffin 1:20-1:50
  • Immunoprecipitation 1:10-1:500
  • In vitro assay reported in multiple pieces of scientific literature
  • Knockdown Validated reported in scientific literature (PMID 32772041)
  • Knockout Validated reported in scientific literature (PMID 26861754)
  • Ligand Activation reported in scientific literature (PMID 26147748)
  • Proximity Ligation Assay reported in scientific literature (PMID 27595394)
  • Simple Western
  • Tissue Culture Substratum
  • Western Blot 1:500
• Application Notes: In WB, a band can be seen at 120 kDa representing HIF-1 alpha in induced tissues and cells. Multiple bands may be seen at 100-120 kDa representing post-translational modification of HIF-1 alpha. For WB, testing on nuclear extracts is recommended. We recommend the use of a highly sensitive ECL reagent, such as West Pico PLUS, for Western blot detection. See Simple Western Antibody Database for Simple Western validation: separated by size
• Theoretical MW: 93 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  COS-7 Nuclear Hypoxic Induced Cell Lysate
  HeLa CoCl2 treated/Untreated Cell Lysate
  HepG2 CoCl2 treated/Untreated Cell Lysate
  HeLa Hypoxic / Normoxic Cell Lysate
  HepG2 Hypoxic / Normoxic Cell Lysate
  HIF-1 alpha Knockout CoCl2-treated/untreated HeLa Cell Lysate
  HIF-1 alpha Knockout Hypoxic-treated/untreated HeLa Cell Lysate
  NIH 3T3 CoCl2 treated/Untreated Cell Lysate
  L929 CoCl2 treated/Untreated Cell Lysate
  Raw 264.7 CoCl2 treated/Untreated Cell Lysate
  PC12 CoCl2 treated/Untreated Cell Lysate
  PC12 Hypoxic / Normoxic Cell Lysate
  Raw 264.7 Hypoxic / Normoxic Cell Lysate
  L929 Hypoxic / Normoxic Cell Lysate
  NIH 3T3 Hypoxic / Normoxic Cell Lysate

Reactivity Notes

Use in Rat reported in scientific literature (PMID:33816617).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS, 1% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for HIF-1 alpha Antibody (H1alpha67)

• AINT
• anti-HIF-1 alpha
• anti-HIF1A
• ARNT interacting protein
• ARNT-interacting protein
• Basic-helix-loop-helix-PAS protein MOP1
• BHLHE78
• Class E basic helix-loop-helix protein 78
• H1alpha67
• HIF 1A
• HIF1 alpha
• HIF-1 alpha
• HIF1
• HIF1A
• HIF-1a
• HIF-1alpha
• HIF-1-alpha
• HIF1-alpha
• hypoxia inducible factor 1 alpha subunit, hypoxia inducible factor 1 subunit alpha
• hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
• hypoxia-inducible factor 1-alpha
• Member of PAS protein 1
• member of PAS superfamily 1
• MOP1
• PAS domain-containing protein 8
• PASD8

Background

Hypoxia contributes to the pathophysiology of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease (1). In cancer and particularly solid tumors, hypoxia plays a critical role in the regulation of genes involved in stem cell renewal, epithelial to mesenchymal transition (EMT), metastasis and angiogenesis. In the tumor microenvironment (TME), hypoxia influences the properties and function of stromal cells (e.g., fibroblasts, endothelial and immune cells) and is a strong determinant of tumor progression (2,3).

HIF-1 or hypoxia inducible factor 1 (predicted molecular weight 93kDa), is a transcription factor commonly referred to as a "master regulator of the hypoxic response" for its central role in the regulation of cellular adaptations to hypoxia. In its active form under hypoxic conditions, HIF-1 is stabilized by the formation of a heterodimer of HIF-1 alpha and ARNT/HIF-1 beta subunits. Nuclear HIF-1 engages p300/CBP for binding to hypoxic response elements (HREs). This process induces transcription and regulation of genes including EPO, VEGF, iNOS2, ANGPT1 and OCT4 (4,5).

Under normoxic conditions, the HIF-1 alpha subunit is rapidly targeted and degraded by the ubiquitin proteasome system. This process is mediated by prolyl hydroxylase domain enzymes (PHDs), which catalyze the hydroxylation of key proline residues (Pro-402 and Pro-564) within the oxygen-dependent degradation domain of HIF-1 alpha. Once hydroxylated, HIF-1 alpha binds the von Hippel-Lindau tumor suppressor protein (pVHL) for subsequent ubiquitination and proteasomal degradation (4). pVHL dependent regulation of HIF-1 alpha plays a role in normal physiology and disease states. Regulation of HIF-1 alpha by pVHL is critical for the suppressive function of FoxP3+ regulatory Tcells (6). Repression of pVHL expression in chronic lymphocytic leukemia (CLL) B cells leads to HIF-1 alpha stabilization and increased VEGF secretion (7).

References

1. Semenza, G. L., Agani, F., Feldser, D., Iyer, N., Kotch, L., Laughner, E., & Yu, A. (2000). Hypoxia, HIF-1, and the pathophysiology of common human diseases. Advances in Experimental Medicine and Biology.

2. Muz, B., de la Puente, P., Azab, F., & Azab, A. K. (2015). The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia. https://doi.org/10.2147/hp.s93413

3. Huang, Y., Lin, D., & Taniguchi, C. M. (2017). Hypoxia inducible factor (HIF) in the tumor microenvironment: friend or foe? Science China Life Sciences. https://doi.org/10.1007/s11427-017-9178-y

4. Koyasu, S., Kobayashi, M., Goto, Y., Hiraoka, M., & Harada, H. (2018). Regulatory mechanisms of hypoxia-inducible factor 1 activity: Two decades of knowledge. Cancer Science. https://doi.org/10.1111/cas.13483

5. Dengler, V. L., Galbraith, M. D., & Espinosa, J. M. (2014). Transcriptional regulation by hypoxia inducible factors. Critical Reviews in Biochemistry and Molecular Biology. https://doi.org/10.3109/10409238.2013.838205

6. Lee, J. H., Elly, C., Park, Y., & Liu, Y. C. (2015). E3Ubiquitin Ligase VHL Regulates Hypoxia-Inducible Factor-1 alpha to Maintain Regulatory T Cell Stability and Suppressive Capacity. Immunity. https://doi.org/10.1016/j.immuni.2015.05.016

7. Ghosh, A. K., Shanafelt, T. D., Cimmino, A., Taccioli, C., Volinia, S., Liu, C. G., ... Kay, N. E. (2009). Aberrant regulation of pVHL levels by microRNA promotes the HIF/VEGF axis in CLL B cells. Blood. https://doi.org/10.1182/blood-2008-10-185686

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HIF-1 alpha Antibody (NB100-105)(1037)

Publications using NB100-105

• Jia Y, Li R, Li Y et Al. Melanoma bone metastasis-induced osteocyte ferroptosis via the HIF1?-HMOX1 axis Bone Res 2025-01-16 [PMID: 39814705]
• Garcia MJ, Morales MS, Yang TS et al. Adverse effects of CXCR2 deficiency in mice reared under non-gnotobiotic conditions Scientific Reports 2024-10-30 [PMID: 39478033]
• Xu Y, Chang L, Chen Y et Al. USP26 Combats Age-Related Declines in Self-Renewal and Multipotent Differentiation of BMSC by Maintaining Mitochondrial Homeostasis Adv Sci (Weinh) 2024-10-08 [PMID: 39377219]
• Raval K, Jamshidi N, Seyran B et Al. Dysfunctional ?-cell longevity in diabetes relies on energy conservation and positive epistasis Life Sci Alliance 2024-09-23 [PMID: 39313296]
• Liu Z, Zheng J, Ding T et Al. HIF-1? protects nucleus pulposus cells from oxidative stress-induced mitochondrial impairment through PDK-1 Free Radic Biol Med 2024-08-10 [PMID: 39128487]
• Jiao M, Hu M, Pan D, Liu X et Al. VHL loss enhances antitumor immunity by activating the anti-viral DNA-sensing pathway iScience 2024-07-25 [PMID: 39050705]
• Rezaei Adariani S, Agne D, Koska S et Al. Detection of a Mitochondrial Fragmentation and Integrated Stress Response Using the Cell Painting Assay J Med Chem 2024-08-08 [PMID: 39018123]
• Qiu, M;Chang, L;Tang, G;Ye, W;Xu, Y;Tulufu, N;Dan, Z;Qi, J;Deng, L;Li, C; Activation of the osteoblastic HIF-1? pathway partially alleviates the symptoms of STZ-induced type 1 diabetes mellitus via RegIII? Experimental & molecular medicine 2024-07-01 [PMID: 38945950]
• Prieto-Vila M, Usuba W, Yoshioka Y et al. High-grade bladder cancer cells secrete extracellular vesicles containing miRNA-146a-5p and promotes angiogenesis. Journal of extracellular biology 2022-08-01 [PMID: 38939052]
• Han H, Kim JE, Lee HJ. et Al. Effect of apigetrin in pseudo-SARS-CoV-2-induced inflammatory and pulmonary fibrosis in vitro model Sci Rep 2024-06-24 [PMID: 38914619]
• Ashutosh Singh, Donghang Cheng, Jyothishmathi Swaminathan, Yanwen Yang, Yan Zheng, Nancy Gordon, Vidya Gopalakrishnan REST-dependent downregulation of von Hippel-Lindau tumor suppressor promotes autophagy in SHH-medulloblastoma Scientific Reports 2024-06-12 [PMID: 38866867]
• Duan M, Ru X, Zhou J et Al. Endothelial EGLN3-PKM2 signaling induces the formation of acute astrocytic barrier to alleviate immune cell infiltration after subarachnoid hemorrhage Fluids Barriers CNS 2024-05-16 [PMID: 38755642]
• Bai W, Ren JS, Li KR et Al. An integrated analysis revealing the angiogenic function of TP53I11 in tumor microenvironment Heliyon 2024-04-15 [PMID: 38655349]
• Li L, Liu Y, Zhi N et Al. Hypoxic preconditioning accelerates the healing of ischemic intestinal injury by activating HIF-1?/PPAR? pathway-mediated fatty acid oxidation Cell Death Discov 2024-04-04 [PMID: 38575595]
• Nathan K. P. Wong, Emma L. Solly, Richard Le, Victoria A. Nankivell, Jocelyne Mulangala, Peter J. Psaltis, Stephen J. Nicholls, Martin K. C. Ng, Christina A. Bursill, Joanne T. M. Tan, Riccardo Alessandro TRIM2 Selectively Regulates Inflammation-Driven Pathological Angiogenesis without Affecting Physiological Hypoxia-Mediated Angiogenesis International Journal of Molecular Sciences 2024-03-15 [PMID: 38542330]
• Fujimoto TN, Colbert LE, Molkentine JM, Baseler L. EGLN Inhibition Reduces Gastrointestinal Radiation Toxicity and Improves Survival in a Murine Model of Locally Advanced Pancreatic Cancer bioRxiv 2018-07-20 (WB, Mouse)
• Nicholas D Nolan, Xuan Cui, Brian M Robbings, Aykut Demirkol, Kriti Pandey, Wen-Hsuan Wu, Hannah F Hu, Laura A Jenny, Chyuan-Sheng Lin, Daniel T Hass, Jianhai Du, James B Hurley, Stephen H Tsang CRISPR editing of anti-anemia drug target rescues independent preclinical models of retinitis pigmentosa. Cell reports. Medicine 2024-04-19 [PMID: 38518771]
• Wen Li, Yongqi Chen, Zhenguo Cai, Xiang He, Lili Yang, Jiong Zhu, Wuqing Wang Traditional Chinese medicine Qingre Huoxue decoction enhances wound healing in through modulation of angiogenic and inflammatory pathways International Wound Journal 2024-03-04 [PMID: 38439195]
• Weaver J, Casellini C, Parson H, Vinik A. EXPRESSION OF HIF-1a IN A PANCREATIC DUCTAL ADENOCARCINOMA IN A PATIENT WITH NEWLY DIAGNOSED TYPE 2 DIABETES AACE Clinical Case Reports 2018-05-01
• Shaun Haran, Kantaraja Chindera, May Sabry, Nafisa Wilkinson, Rupali Arora, Agnieszka Zubiak, Thomas E. Bartlett, Iona Evans, Allison Jones, Daniel Reisel, Chiara Herzog, Twana Alkasalias, Mark Newman, Jaeyeon Kim, Angelique Flöter Rådestad, Kristina Gemzell-Danielsson, Adam N. Rosenthal, Louis Dubeau, Mark W. Lowdell, Martin Widschwendter, Ugo Cavallaro Natural Killer Cell Dysfunction in Premenopausal BRCA1 Mutation Carriers: A Potential Mechanism for Ovarian Carcinogenesis Cancers 2024-03-18 [PMID: 38539519]
• Shiu-Ju Yang, Chih-Hao Hsu, Chi-Yun Lai, Pei-Chu Tsai, Yung-Deng Song, Chang-Ching Yeh, Yih-Yuan Chen, Horng-Yunn Dou Pathological granuloma fibrosis induced by agar-embedded Mycobacterium abscessus in C57BL/6JNarl mice Frontiers in Immunology 2023-12-11 [PMID: 38146374]
• Manivannan Yegambaram, Xutong Sun, Alejandro Garcia Flores, Qing Lu, Jamie Soto, Jaime Richards, Saurabh Aggarwal, Ting Wang, Haiwei Gu, Jeffrey R. Fineman, Stephen M. Black, Venediktova Natalya Novel Relationship between Mitofusin 2-Mediated Mitochondrial Hyperfusion, Metabolic Remodeling, and Glycolysis in Pulmonary Arterial Endothelial Cells International Journal of Molecular Sciences 2023-12-15 [PMID: 38139362]
• Pan YD, Zhang Y, Zheng WY et al. Intermittent Hypobaric Hypoxia Ameliorates Autistic-Like Phenotypes in Mice. The Journal of neuroscience : the official journal of the Society for Neuroscience 2024-02-16 [PMID: 38124211]
• Niu N, Miao H, Ren H Effect of miR-182-5p on apoptosis in myocardial infarction Heliyon 2023-10-01 [PMID: 38034598] (WB, Rat)
  
  Details:
  1:1000 WB dilution
• Liu Z, Geng Y, Huang Y et al. Bushen Antai Recipe Alleviates Embryo Absorption by Enhancing Immune Tolerance and Angiogenesis at Maternal-Fetal Interface Via Mobilizing Mdscs in Abortion-Prone Mice SSRN 2023-06-24 [PMID: 37952407] (WB, Mouse)
• Cimmino TP, Pagano E, Stornaiuolo M et al. Formyl-peptide receptor 2 signalling triggers aerobic metabolism of glucose through Nox2-dependent modulation of pyruvate dehydrogenase activity Open biology 2023-10-01 [PMID: 37875162] (WB, Human)
• Soeda K, Sasako T, Enooku K et al. Gut insulin action protects from hepatocarcinogenesis in diabetic mice comorbid with nonalcoholic steatohepatitis Nature communications 2023-10-18 [PMID: 37852976] (WB, Mouse)
• Yilmaz-Oral D, Sezen SF, Turkcan D et al. Dual Strategy with Adipose-derived Stem Cells and L-arginine Recovered Cavernosal Functions in a Rat Model of Radical Prostatectomy Stem cells and development 2023-10-17 [PMID: 37847152]
• Qiang Zhou, Wei Huang, Jing Xiong, Biao Guo, Xinghuan Wang, Ju Guo CDCA8 promotes bladder cancer survival by stabilizing HIF1 alpha expression under hypoxia Cell Death & Disease 2023-10-09 [PMID: 37813876]
• Joo HY, Jung JK, Kim MY et al. NADH elevation during chronic hypoxia leads to VHL-mediated HIF-1? degradation via SIRT1 inhibition Cell & bioscience 2023-09-30 [PMID: 37777750] (IP, Human)
• Shirai T, Uemichi K, Takemasa T Effects of the order of endurance and high intensity interval exercise in combined training on mice skeletal muscle metabolism American journal of physiology. Regulatory, integrative and comparative physiology 2023-09-25 [PMID: 37746708]
• Suresh MV, Aktay S, Yalamanchili G et al. Role of succinate in airway epithelial cell regulation following traumatic lung injury JCI insight 2023-09-22 [PMID: 37737265] (Simple Western, Mouse)
  
  Details:
  1:50 dilution
• Schoelles KJ, Kemper K, Martin G et al. HIF1? and HIF2? immunoreactivity in epithelial tissue of primary and recurrent pterygium by immunohistochemical analysis International ophthalmology 2023-09-08 [PMID: 37684398] (IHC-P, Human)
• Inada A, Fukatsu A Persistence and expansion of hypoxia detected by pimonidazole-adduct immunostaining during progression of diabetic nephropathy in diabetic mice American journal of physiology. Renal physiology 2023-08-24 [PMID: 37615048]
• Xie Y, Guo W, Shen X et al. A delayed ovulation of progestin-primed ovarian stimulation (PPOS) by downregulating the LHCGR/PGR pathway iScience 2023-08-18 [PMID: 37520702] (ICC/IF, Mouse)
• Liu Z, Song Y, Hu R et al. Bushen Antai recipe ameliorates immune microenvironment and maternal-fetal vascularization in STAT3-deficient abortion-prone mice Journal of ethnopharmacology 2023-07-08 [PMID: 37423519] (WB, Mouse)
• Oreši? T, Bubanovi? S, Rami? S et al. Nuclear localization of NRF2 in stroma of HER2 positive and triple-negative breast cancer Pathology, research and practice 2023-07-05 [PMID: 37421843] (IHC-P, Human)
• Zhang M, Shi Z, Peng X et al. NLRP3 inflammasome-mediated Pyroptosis induce Notch signal activation in endometriosis angiogenesis Molecular and cellular endocrinology 2023-05-31 [PMID: 37268099] (Western Blot, Immunocytochemistry/ Immunofluorescence, Human, Mouse)
  
  Details:
  ICC/IF 1:200, WB 1:1000
• Zhang L, Xu S, Cheng X et al. Inflammatory tumor microenvironment of thyroid cancer promotes cellular dedifferentiation and silencing of iodide-handling genes expression Pathology - Research and Practice 2023-05-01 [PMID: 37172523]
• Chang E, Wang Y, Zhu R et al. General anesthetic action profile on the human prefrontal cortex cells through comprehensive single-cell RNA-seq analysis iScience 2023-03-01 [PMID: 37123239] (SDS-Page)
• Zhang H, Wang L, Cui J et al. Maintaining hypoxia environment of subchondral bone alleviates osteoarthritis progression Science advances 2023-04-05 [PMID: 37018403] (IHC-P, Mouse)
• He S, Fan C, Ji Y et al. SENP3 facilitates M1 macrophage polarization via the HIF-1?/PKM2 axis in lipopolysaccharide-induced acute lung injury Innate immunity 2023-04-05 [PMID: 37016838] (WB, Mouse)
• Qiu M, Li C, Cai Z et al. 3D Biomimetic Calcified Cartilaginous Callus that Induces Type H Vessels Formation and Osteoclastogenesis Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2023-03-31 [PMID: 36999832] (IHC-P, Rat)
• Gambardella J, Fiordelisi A, Cerasuolo FA, Buonaiuto A et Al. Experimental evidence and clinical implications of Warburg effect in the skeletal muscle of Fabry disease iScience 2023-03-07 [PMID: 36879801]
• Hernendez-Rollan N, Sanchez-Mateos S, Lopez-Morato M et al. Supplementation of culture medium with quercetin improves mouse blastocyst quality and increases the expression of HIF-1 alpha protein Zygote (Cambridge, England) 2023-02-27 [PMID: 36843100]
• Wu K, Li B, Ma Y et al. Nicotinamide mononucleotide attenuates HIF-1? activation and fibrosis in hypoxic adipose tissue via NAD+/SIRT1 axis Frontiers in endocrinology 2023-01-26 [PMID: 36777361] (Immunoprecipitation, Mouse)
• Wu K, Wu M, Yang H et al. Hypoxia promotes conversion to a stem cell phenotype in prostate cancer cells by activating HIF-1?/Notch1 signaling pathway Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico 2023-02-09 [PMID: 36757381]
• Leca J, Lemonnier F, Meydan C et al. IDH2 and TET2 mutations synergize to modulate T Follicular Helper cell functional interaction with the AITL microenvironment Cancer Cell 2023-02-01 [PMID: 36736318] (IHC-P, Human)
• Wang C, Wu Z, Li Z et al. Beneficial effect of the mitochondrial ATP-sensitive potassium channel-specific opener nicorandil on the collapsed lung via inhibition of apoptosis in clinical thoracic surgery Molecular medicine reports 2023-03-01 [PMID: 36734261]
• Kong Y, Liu PK, Li Y et al. HIF2? activation and mitochondrial deficit due to iron chelation cause retinal atrophy EMBO molecular medicine 2023-02-08 [PMID: 36645044] (Western Blot)
• Sieow J, Penny H, Gun S et al. Conditional Knockout of Hypoxia-Inducible Factor 1-Alpha in Tumor-Infiltrating Neutrophils Protects against Pancreatic Ductal Adenocarcinoma International Journal of Molecular Sciences 2023-01-01 [PMID: 36614196] (IHC-P, Human, Mouse)
  
  Details:
  Dilution used in IHC-P 1:100
• Liu C, Meng M, Xu B et al. Fgf6 Promotes Adipocyte Progenitor Cell Proliferation for Adipose Tissue Homeostasis SSRN Electronic Journal 2022-03-19 [PMID: 36607240]
• Park SJ, Yoo HC, Ahn E, Luo E et Al. Enhanced Glutaminolysis Drives Hypoxia-Induced Chemoresistance in Pancreatic Cancer Cancer Res 2023-01-03 [PMID: 36594876]
• Zhong WJ, Liu T, Yang HH et al. TREM-1 governs NLRP3 inflammasome activation of macrophages by firing up glycolysis in acute lung injury International journal of biological sciences 2023-01-01 [PMID: 36594089] (ICC/IF, Mouse)
  
  Details:
  Dilution used in ICC 1:100
• David BT, Curtin JJ, Brown JL et al. Temporary induction of hypoxic adaptations by preconditioning fails to enhance Schwann cell transplant survival after spinal cord injury Glia 2022-12-24 [PMID: 36565279]
• Chen J, Zhang M, Liu Y et al. Histone lactylation driven by mROS-mediated glycolytic shift promotes hypoxic pulmonary hypertension Journal of molecular cell biology 2022-12-23 [PMID: 36564027] (ICC/IF, IP, IHC-P, Rat)
  
  Details:
  Dilution used for IHC-P 1:50
• Tang T, Jia Y, Liang H et al. Knockdown of Nrf2 radiosensitizes glioma cells by inducing redox stress and apoptosis in hypoxia Translational Cancer Research 2022-11-01 [PMID: 36523295] (Immunohistochemistry)
• DAtilo MN, Formigari GP, de Faria JBL, de Faria JML AMP kinase activation by Omega-3 polyunsaturated fatty acid protects the retina against ischemic insult: An in vitro and in vivo study Experimental eye research 2022-12-09 [PMID: 36509164] (ICC/IF, IHC, Mouse, Human)
• Dai T, Rosario SR, Katsuta E et al. Hypoxic activation of PFKFB4 in breast tumor microenvironment shapes metabolic and cellular plasticity to accentuate metastatic competence Cell reports 2022-12-06 [PMID: 36476868]
• Yang H, Wu Y, Cheng M et al. Roxadustat (FG-4592) protects against ischemia-induced acute kidney injury via improving CD73 and decreasing AIM2 inflammasome activation Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association 2022-11-22 [PMID: 36413468]
• Zhang Y, Song Y, Ren S et al. GPER-mediated stabilization of HIF-1 alpha contributes to upregulated aerobic glycolysis in tamoxifen-resistant cells Oncogene 2022-11-18 [PMID: 36400971] (IF/IHC, Human)
• He YL, Li J, Gong SH et al. BNIP3 phosphorylation by JNK1/2 promotes mitophagy via enhancing its stability under hypoxia Cell death & disease 2022-11-17 [PMID: 36396625] (WB, Rat)
  
  Details:
  Dilution used in WB 1:1000
• Tang J, Wu Z, Wang X et al. Hypoxia-Regulated lncRNA USP2-AS1 Drives Head and Neck Squamous Cell Carcinoma Progression Cells 2022-10-28 [PMID: 36359803] (WB, Chemotaxis, Human)
• Vohwinkel CU, Burns N, Coit E et al. HIF1A-dependent induction of alveolar epithelial PFKFB3 dampens acute lung injury JCI insight 2022-11-03 [PMID: 36326834] (WB, Mouse)
• Ji Y, Liang Y, Chu PH Et al. The effects of intermittent hypoxia on hepatic expression of fatty acid translocase CD36 in lean and diet-induced obese mice Biomed J 2022-10-16 [PMID: 36244649] (IHC-P, Mouse)
  
  Details:
  Citation using the HRP version of this antibody.
• Li D, Wang T, Lai J et al. Silencing TRPM2 enhanced erastin- and RSL3-induced ferroptosis in gastric cancer cells through destabilizing HIF-1 alpha and Nrf2 proteins Cytotechnology 2022-08-27 [PMID: 36238268]
• Rosa P, Scibetta S, Pepe G et al. Polysialic Acid Sustains the Hypoxia-Induced Migration and Undifferentiated State of Human Glioblastoma Cells International Journal of Molecular Sciences 2022-08-24 [PMID: 36076963] (WB, IHC-P, Human)
  
  Details:
  WB dilution used 1:1000, IHC-P dilution 1:100
• Huang YJ, Chen YT, Huang CM et al. HIF-1 alpha Expression Increases Preoperative Concurrent Chemoradiotherapy Resistance in Hyperglycemic Rectal Cancer Cancers 2022-08-22 [PMID: 36011045] (IHC-P, Human)
  
  Details:
  Dilution used 1:500
• Igbo BT, Linge A, Frosch S et al. Immunohistochemical analyses of paraffin-embedded sections after primary surgery or trimodality treatment in esophageal carcinoma Clinical and translational radiation oncology 2022-09-01 [PMID: 35993091] (IHC-P, Human)
• Wang K, Chen YS, Chien HW et al. Melatonin inhibits NaIO3-induced ARPE-19 cell apoptosis via suppression of HIF-1 alpha/BNIP3-LC3B/mitophagy signaling Cell & bioscience 2022-08-19 [PMID: 35986432] (ICC/IF, WB, Chemotaxis, KD, IF/IHC, Human, Mouse)
• Wang P, Zeng G, Yan Y Et al. Disruption of adipocyte HIF-1 alpha improves atherosclerosis through the inhibition of ceramide generation Acta Pharm Sin B 2022-07-18 [PMID: 35847503]
• Wang X, Zhang X, Cao K et al. Cardiac disruption of SDHAF4-mediated mitochondrial complex II assembly promotes dilated cardiomyopathy Nature communications 2022-07-08 [PMID: 35803927] (IB, Mouse)
  
  Details:
  Dilution: 1:2000
• Reiji Ito, Elizabeth A Barnes, Xibing Che, Cristina M Alvira, David N Cornfield SM22 alpha cell-specific HIF stabilization mitigates hyperoxia-induced neonatal lung injury. American journal of physiology. Lung cellular and molecular physiology 2022-08-02 [PMID: 35762602]
• Qin Y, Dinabandhu A, Cao X et al. ANGPTL4 influences the therapeutic response of neovascular age-related macular degeneration patients by promoting choroidal neovascularization JCI insight 2022-06-02 [PMID: 35653189] (IF/IHC, Mouse)
• He WZ, Yang M, Jiang Y et al. miR-188-3p targets skeletal endothelium coupling of angiogenesis and osteogenesis during ageing Cell death & disease 2022-05-25 [PMID: 35610206] (WB, Mouse)
• Nan Niu, Hui Li, Xiancai Du, Chan Wang, Junliang Li, Jihui Yang, Cheng Liu, Songhao Yang, Yazhou Zhu, Wei Zhao Effects of NRF-1 and PGC-1 alpha cooperation on HIF-1 alpha and rat cardiomyocyte apoptosis under hypoxia. Gene 2022-06-21 [PMID: 35569770]
• Jacquier V, Gitenay D, Fritsch S et al. RIP140 inhibits glycolysis-dependent proliferation of breast cancer cells by regulating GLUT3 expression through transcriptional crosstalk between hypoxia induced factor and p53 Cellular and molecular life sciences : CMLS 2022-05-03 [PMID: 35501580] (WB, Mouse)
  
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  Fig. 1
• RytkOnen K, Faux T, Mahmoudian M et al. Histone H3K4me3 breadth in hypoxia reveals endometrial core functions and stress adaptation linked to endometriosis iScience 2022-05-01 [PMID: 35494227]
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• Inada A, Inada O, Yasunami Y et al. Amelioration of murine diabetic nephropathy with a SGLT2 inhibitor is associated with suppressing abnormal expression of hypoxia-inducible factors The American journal of pathology 2022-04-20 [PMID: 35460614] (IF/IHC, Mouse)
• Xia QS, Wu F, Wu WB et al. Berberine reduces hepatic ceramide levels to improve insulin resistance in HFD-fed mice by inhibiting HIF-2 alpha Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 2022-04-13 [PMID: 35429745] (IHC-P, WB, ICC/IF, Human, Mouse)
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• Min, J;Ma, F;Seyran, B;Pellegrini, M;Greeff, O;Moncada, S;Tudzarova, S; beta-cell-specific deletion of PFKFB3 restores cell fitness competition and physiological replication under diabetogenic stress Communications biology [PMID: 35318430] (ICC/IF, Mouse)
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• Chen TH, Liu CT, Cheng Cy et al. Oligosaccharides Ameliorate Acute Kidney Injury by Alleviating Cluster of Differentiation 44-Mediated Immune Responses in Renal Tubular Cells Nutrients 2022-02-11 [PMID: 35215410]
• Liu Y, Cong Y, Niu Y et al. Themis is indispensable for IL-2 and IL-15 signaling in T cells Science signaling 2022-02-15 [PMID: 35167340]
• Wei H, Xu Z, Chen L et al. Long non-coding RNA PAARH promotes hepatocellular carcinoma progression and angiogenesis via upregulating HOTTIP and activating HIF-1 alpha/VEGF signaling Cell death & disease 2022-02-02 [PMID: 35110549] (WB, Chemotaxis, Human)
• Burak Kandilci H, Simsek G, Zirapli T, Celebier M Differential role of prolyl hydroxylases on mitochondrial function of HL-1 cells in a model of iron deficiency Mitochondrion 2022-01-20 [PMID: 35066139] (ICC/IF, Mouse)
• Dong X, Wu P, Yan L Et al. Oriented nanofibrous P(MMD-co-LA)/Deferoxamine nerve scaffold facilitates peripheral nerve regeneration by regulating macrophage phenotype and revascularization Biomaterials 2021-12-01 [PMID: 34894585] (IHC-P, Rat)
• Li H, Wang M, Huang B et al. Theranostic near-infrared-IIb emitting nanoprobes for promoting immunogenic radiotherapy and abscopal effects against cancer metastasis Nature communications 2021-12-09 [PMID: 34887404] (WB, IF/IHC, Mouse)
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• Dong W, Song Z, Liu S Et Al. Adipose-Derived Stem Cells Based on Electrospun Biomimetic Scaffold Mediated Endothelial Differentiation Facilitating Regeneration and Repair of Abdominal Wall Defects via HIF-1 alpha/VEGF Pathway Frontiers in bioengineering and biotechnology 2021-07-07 [PMID: 34307320] (ICC/IF)
• Zhou F, Song Y, Liu X Et Al. Si-Wu-Tang facilitates ovarian function through improving ovarian microenvironment and angiogenesis in a mouse model of premature ovarian failure Journal of ethnopharmacology 2021-07-19 [PMID: 34293457] (WB)
• Kumar Singh R, Pei Y, Bose D Et Al. KSHV-encoded vCyclin can modulate HIF1 alpha levels to promote DNA replication in hypoxia eLife 2021-07-19 [PMID: 34279223] (ICC/IF, WB)
• Chen Mh, Wang Yh, Sun Bj Et Al. HIF-1 alpha activator DMOG inhibits alveolar bone resorption in murine periodontitis by regulating macrophage polarization International immunopharmacology 2021-07-14 [PMID: 34273637] (IF/IHC)
• Yu S, Li Q, Wang Y Et Al. Tumor-derived LIF promotes chemoresistance via activating tumor-associated macrophages in gastric cancers Experimental cell research 2021-07-13 [PMID: 34265288] (Chemotaxis, WB)
• Wang Y, Wang X, Wang H Et Al. PTEN protects kidney against acute kidney injury by alleviating apoptosis and promoting autophagy via regulating HIF1-alpha and mTOR through PI3K/Akt pathway Experimental cell research 2021-07-07 [PMID: 34242625] (WB)
• Penny H, Sieow J, Gun S et al. Targeting Glycolysis in Macrophages Confers Protection Against Pancreatic Ductal Adenocarcinoma International Journal of Molecular Sciences 2021-06-14 [PMID: 34198548] (Mouse)
• Nakahara M, Ito H, Skinner Jt Et Al. The inflammatory role of dysregulated IRS2 in pulmonary vascular remodeling under hypoxic conditions American journal of physiology. Lung cellular and molecular physiology 2021-06-30 [PMID: 34189964]
• Guo Xh, Wu My, Zhao G Et Al. The locoregional adiponectin and its synergistic antitumor effect with HIF-1alpha blockade in TSCC Oral diseases 2021-06-26 [PMID: 34174132]
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• Hirayama Y, Anzai N,Koizumi S Mechanisms underlying sensitization of P2X7 receptors in astrocytes for induction of ischemic tolerance Glia 2021-06-02 [PMID: 34076906]
• Yao X, Yang W, Ren Z et al. Neuroprotective and Angiogenesis Effects of Levetiracetam Following Ischemic Stroke in Rats Frontiers in pharmacology 2021-05-14 [PMID: 34054520] (WB, Rat)
• David BT, Curtin JJ, Brown JL et al. Treatment with hypoxia-mimetics protects cultured rat Schwann cells against oxidative stress-induced cell death Glia 2021-05-21 [PMID: 34019306]
• Paul S, Gangwar A, Arya A et al. Modulation of lung cytoskeletal remodeling, RXR based metabolic cascades and inflammation to achieve redox homeostasis during extended exposures to lowered pO2 Apoptosis : an international journal on programmed cell death 2021-05-17 [PMID: 34002323] (WB, Mouse, Human)
• Shang FF, Wang JY, Xu Q et al. Design, synthesis of novel celastrol derivatives and study on their antitumor growth through HIF-1 alpha pathway European journal of medicinal chemistry 2021-04-21 [PMID: 33930802]
• Favara DM, Liebscher I, Jazayeri A et al. Elevated expression of the adhesion GPCR ADGRL4/ELTD1 promotes endothelial sprouting angiogenesis without activating canonical GPCR signalling Scientific reports 2021-04-23 [PMID: 33893326] (WB, Human)
• Wang M, Li H, Huang B et al. An Ultra-Stable, Oxygen-Supply Nanoprobe Emitting in Near-Infrared-II Window to Guide and Enhance Radiotherapy by Promoting Anti-Tumor Immunity Advanced healthcare materials 2021-04-22 [PMID: 33885213]
• Kuwano A, Kurokawa M, Kohjima M et al. Microcirculatory disturbance in acute liver injury Experimental and therapeutic medicine 2021-06-01 [PMID: 33884034] (IF/IHC, Human)
• Yoshikawa M, Morine Y, Yamada S et al. Prognostic prediction of resectable colorectal liver metastasis using the apparent diffusion coefficient from diffusion weighted magnetic resonance imaging Ann Gastroenterol Surg 2021-04-16 [PMID: 33860146]
• Wang C, Niu F, Ren N et al. Hyperbaric Oxygen Improves Cerebral Ischemia/Reperfusion Injury in Rats Probably via Inhibition of Autophagy Triggered by the Downregulation of Hypoxia-Inducing Factor-1 Alpha BioMed Research International 2021-03-15 [PMID: 33816617] (ICC/IF, WB, Rat)
• Zheng J, Chang L, Bao X et al. TRIM21 drives intervertebral disc degeneration induced by oxidative stress via mediating HIF-1alpha degradation Biochemical and Biophysical Research Communications 2021-05-01 [PMID: 33813275] (IHC-P, Mouse)
• SAnchez-Del-Campo L, MartI-DIaz R, Montenegro MF et al. MITF induces escape from innate immunity in melanoma Journal of experimental & clinical cancer research : CR 2021-03-31 [PMID: 33789714] (Human)
• HOper T, Siewert K, Dumit VI et al. The Contact Allergen NiSO4 Triggers a Distinct Molecular Response in Primary Human Dendritic Cells Compared to Bacterial LPS Frontiers in immunology 2021-03-11 [PMID: 33777040] (WB, Human)
• Zeng J, Xu H, Huang C et al. CD46 splice variant enhances translation of specific mRNAs linked to an aggressive tumor cell phenotype in bladder cancer Molecular Therapy - Nucleic Acids 2021-02-01 [PMID: 33767911]
• Vohwinkel CU, Coit EJ, Burns N et al. Targeting alveolar-specific succinate dehydrogenase A attenuates pulmonary inflammation during acute lung injury FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2021-04-01 [PMID: 33687752] (WB, Mouse)
• Sakai T, Matsuo Y, Okuda K et al. Development of antitumor biguanides targeting energy metabolism and stress responses in the tumor microenvironment Scientific reports 2021-03-01 [PMID: 33649449] (WB, Human)
• Fan T, Feng X, Yokota A et al. Arsenic Dispensing Powder Promotes Erythropoiesis in Myelodysplastic Syndromes via Downregulation of HIF1A and Upregulation of GATA Factors The American journal of Chinese medicine 2021-02-25 [PMID: 33641653]
• Wang B, Li J, Wang Y Magnetic resonance diffusion tensor imaging applied to rat model of contrast-induced acute kidney injury PeerJ 2021-02-15 [PMID: 33628631] (IF/IHC, Rat)
• He R, Wang Z, Cui M, et al. HIF1A Alleviates compression-induced apoptosis of nucleus pulposus derived stem cells via upregulating autophagy Autophagy 2021-01-18 [PMID: 33455530] (IF/IHC, Human, Rat)
• Paul S, Gangwar A, Bhargava K, Ahmad Y D4F prophylaxis enables redox and energy homeostasis while preventing inflammation during hypoxia exposure Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 2021-01-01 [PMID: 33378979] (WB, Rat)
• Shi Y, Liu Z, Zhang Q et al. Phosphorylation of seryl-tRNA synthetase by ATM/ATR is essential for hypoxia-induced angiogenesis PLoS biology 2020-12-01 [PMID: 33351793] (WB, IF/IHC, Mouse, Human)
• Q Lin, S Li, N Jiang, H Jin, X Shao, X Zhu, J Wu, M Zhang, Z Zhang, J Shen, W Zhou, L Gu, R Lu, Z Ni Inhibiting NLRP3 inflammasome attenuates apoptosis in contrast-induced acute kidney injury through the upregulation of HIF1A and BNIP3-mediated mitophagy Autophagy, 2020-12-19;0(0):1-16. 2020-12-19 [PMID: 33345685]
• Wan X, Xie MK, Xu H et al. Hypoxia-preconditioned adipose-derived stem cells combined with scaffold promote urethral reconstruction by upregulation of angiogenesis and glycolysis Stem cell research & therapy 2020-12-11 [PMID: 33308306] (WB, Rabbit, Human)
• Xie H, Xu G, Gao Y, Yuan Z hCINAP serves a critical role in hypoxia induced cardiomyocyte apoptosis via modulating lactate production and mitochondrial mediated apoptosis signaling Molecular medicine reports 2021-02-01 [PMID: 33300073] (WB, Chemotaxis, Human)
• Alijani-Ghazyani Z, Roushandeh AM, Sabzevari R et al. Conditioned medium harvested from Hif1alpha engineered mesenchymal stem cells ameliorates LAD-occlusion -induced injury in rat acute myocardial ischemia model The international journal of biochemistry & cell biology 2021-01-01 [PMID: 33279679] (WB, Human)
• Singh A, Chow O, Jenkins S et al. Characterizing Ischaemic Tolerance in Rat Pheochromocytoma (PC12) Cells and Primary Rat Neurons Neuroscience 2020-11-24 [PMID: 33246056] (Rat)
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• Gauthier LR, Saati M, Bensalah-Pigeon H et al. The HIF1 alpha/JMY pathway promotes glioblastoma stem-like cell invasiveness after irradiation Sci Rep 2020-10-30 [PMID: 33128011] (IF/IHC, Mouse)
• Moon Y, Kim I, Chang S et al. Sigma-1 receptor regulates mitophagy in dopaminergic neurons and contributes to dopaminergic protection Biochim Biophys Acta Gene Regul Mech 2020-10-26 [PMID: 33035707] (Chemotaxis, Human)
• Ganji N Remote Ischemic Conditioning during Experimental Necrotizing Enterocolitis: Mechanism of Action Thesis 2021-01-01 [PMID: 33009377] (IF/IHC, IF/IHC, Human)
• de Laia AGS, Valverde TM, Barrioni BR et al. Cobalt-containing bioactive glass mimics vascular endothelial growth factor A and hypoxia inducible factor 1 function J Biomed Mater Res A 2020-09-02 [PMID: 32876363]
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• Wang Z, Cui M, Qu Y et al. Hypoxia protects rat bone marrow mesenchymal stem cells against compression-induced apoptosis in degenerative disc microenvironment through activation of the HIF-1 alpha/YAP signaling pathway Stem Cells Dev. 2020-08-16 [PMID: 32799744] (WB)
• Fang X, Zhang D, Zhao W et al. Dishevelled Associated Activator Of Morphogenesis (DAAM) Facilitates Invasion of Hepatocellular Carcinoma by Upregulating Hypoxia-Inducible Factor 1 alpha (HIF-1alpha) Expression Med. Sci. Monit. 2020-08-09 [PMID: 32772041] (KD, WB)
• Valeria Capaci, Lorenzo Bascetta, Marco Fantuz, Galina V. Beznoussenko, Roberta Sommaggio, Valeria Cancila, Andrea Bisso, Elena Campaner, Alexander A. Mironov, Jacek R. Wiśniewski, Luisa Ulloa Severino, Denis Scaini, Fleur Bossi, Jodi Lees, Noa Alon, Ledia Brunga, David Malkin, Silvano Piazza, Licio Collavin, Antonio Rosato, Silvio Bicciato, Claudio Tripodo, Fiamma Mantovani, Giannino Del Sal Mutant p53 induces Golgi tubulo-vesiculation driving a prometastatic secretome Nature Communications 2020-08-07 [PMID: 32770028]
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  Details:
  Citation using the HRP format of this antibody.
• Tran MGB, Bibby BAS, Yang L et al. Independence of HIF1a and androgen signaling pathways in prostate cancer BMC Cancer 2020-05-27 [PMID: 32450824]
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  Details:
  Mouse monoclonal anti-HIF-1 alpha antibody used for IHC on paraffin-embedded livers from untreated and PCB 126 treated rats at a dilution of 1:50, incubated for 2 hours at room temperature (Figure 3-1)
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  Details:
  Citation using the Biotin version of this antibody.
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  Details:
  This citation used the DyLight 550 version of this antibody.
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  Details:
  HIF-2 alpha/EPAS1 was used at 1:500 dilution for WB assays in experiments involving human kidney cell lines namely ACHN, Caki-1 and HEK293T.
• Maugeri G, D'Amico AG, Reitano R et al. Parkin modulates expression of HIF-1alpha and HIF-3alpha during hypoxia in gliobastoma-derived cell lines in vitro. Cell Tissue Res. 2016-01-08 [PMID: 26742768] (ICC/IF, WB, Human)
  
  Details:
  HIF-1 alpha (clone H1alpha67) was used at 1:500 dilution for WB assays in lysates from human glioblastoma cell lines U87MG and T98G which were left untransfected or transfected with empty vector (Control shRNA) or with Parkin shRNA (Parkin shRNA) followed by culturing under normoxia or hypoxia conditions. The antibody was used for immunofluorescence analysis (ICC/IF) also on the same cells under similar experimental conditions.
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• Clancy KW, Weerapana E, Thompson PR Detection and identification of protein citrullination in complex biological systems. Curr Opin Chem Biol 2016-01-01 [PMID: 26517730]
• Cho MH, Park CK, Park M et al. Clinicopathologic Features and Molecular Characteristics of Glucose Metabolism Contributing to 18F-fluorodeoxyglucose Uptake in Gastrointestinal Stromal Tumors. PLoS ONE. 2015-10-29 [PMID: 26509967] (IHC-P, Human)
• Zhang L, Takara K, Yamakawa D et al. Apelin as a marker for monitoring the tumor vessel normalization window during antiangiogenic therapy. Cancer Sci. 2015-10-17 [PMID: 26475217] (WB, Mouse)
• Kwang Kim K, Abelman S, Yano N et al. Tetrathiomolybdate inhibits mitochondrial complex IV and mediates degradation of hypoxia-inducible factor-1a in cancer cells. Sci Rep. 2015-10-16 [PMID: 26469226] (WB, Human)
• Cui H, Qin Q, Yang M et al. Bortezomib enhances the radiosensitivity of hypoxic cervical cancer cells by inhibiting HIF-1a expression. Int J Clin Exp Pathol 2015-08-15 [PMID: 26464645] (WB, Human)
• Bo Kong, Tao Cheng, Weiwei Wu, Ivonne Regel, Susanne Raulefs, Helmut Friess, Mert Erkan, Irene Esposito, Jörg Kleeff, Christoph W. Michalski Hypoxia-induced endoplasmic reticulum stress characterizes a necrotic phenotype of pancreatic cancer Oncotarget 2015-10-13 [PMID: 26452217]
• Abe T, Kitaoka Y, Kikuchi DM et al. High-intensity interval training-induced metabolic adaptation coupled with an increase in Hif-1alpha and glycolytic protein expression. J Appl Physiol (1985) 2015-12-01 [PMID: 26429867]
• Yu MO, Park KJ, Park DH et al. Reactive oxygen species production has a critical role in hypoxia-induced Stat3 activation and angiogenesis in human glioblastoma. J. Neurooncol. 2015-08-22 [PMID: 26297045] (WB, Human)
  
  Details:
  HIF1 alpha antibody was used for WB on lysates of human glioblastoma cell line U87 subjected to growth in vitro under hypoxic conditions (Fig. 1A).
• Donato MD, Fanelli M, Mariani M et al. Nek6 and Hif-1alpha cooperate with the cytoskeletal gateway of drug resistance to drive outcome in serous ovarian cancer. Am J Cancer Res 2015-01-01 [PMID: 26269749] (IP)
• Belo AI, van Vliet SJ, Maus A et al. Hypoxia inducible factor 1alpha down regulates cell surface expression of alpha1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. FEBS Lett. 2015-08-19 [PMID: 26232512] (ICC/IF, Human)
• Wang IK, Sun KT, Tsai TH et al. MiR-20a-5p mediates hypoxia-induced autophagy by targeting ATG16L1 in ischemic kidney injury Life Sci. 2015-07-10 [PMID: 26165754] (WB, Human)
• Veerman KJ. Investigating the Pro-Atherogenic Potential of Chronic Hyperglycemia: Is Diabetic Atherosclerosis a Microvascular Complication? [Internet]. 2011-01-01
• Frey JL, Stonko DP, Faugere MC, Riddle RC Hypoxia-inducible factor-1alpha restricts the anabolic actions of parathyroid hormone. Bone Res 2014-05-13 [PMID: 26273518] (IP)
• Dai T, Rosario S, Katsuta E et al. Hypoxic Tumor Microenvironment Induces Metabolic Enzyme PFKFB4 Acquiring Invasive Cancer Phenotype to Drive Metastasis SSRN Electronic Journal 2022-04-22
• de la Puente P, Muz B, Gilson RC et al 3D tissue-engineered bone marrow as a novel model to study pathophysiology and drug resistance in multiple myeloma. Biomaterials. 2015-12-01 [PMID: 26402156] (IHC-P, Human)
  
  Details:
  Citation using the Alexa Fluor 488 version of this antibody.
• PAUL S, GANGWAR A, ARYA A, Ahmad Y Interplay of lung cytoskeletal re-modeling, energy metabolism and inflammation during extended exposures to lowered pO2 Authorea Preprints Jan 1 2022 (WB, Human)
• Clinch M The Role of Hypoxia on PORCN and WLS Expression in Human Embryonic Kidney (HEK293T) and Human Colon Cancer (HCT-116T) Cells Carleton University 2022-04-05 (WB, Human)
• Singh A Activation of Hypoxia Inducible Factor pathway for neuroprotection in in vitro models of ischaemic stroke Thesis 2022-01-01 (WB, Rat)
• Prieto Vila M, Usuba W, Yoshioka Y et al. High grade bladder cancer cells secrete extracellular vesicles containing miRNA 146a 5p and promotes angiogenesis Journal of Extracellular Biology 2022-08-01 (IHC-P, Mouse)
• Wong CS, Chen A, Liu MC et al. Loss of Siah2 does not impact angiogenic potential of murine endothelial cells. Microvasc. Res. 2015-08-11 [PMID: 26275748] (WB, Mouse)
  
  Details:
  SIAH2 antibody (clone 24E6H3, NB110-88113) was used for WB on lysates of endothelial cells isolated from lungs of wildtype (WT) and Siah2?/? (KO) mice.
• Chan MC, Atasoylu O, Hodson E et al. Potent and Selective Triazole-Based Inhibitors of the Hypoxia-Inducible Factor Prolyl-Hydroxylases with Activity in the Murine Brain PLoS ONE. 2015-07-07 [PMID: 26147748] (IA, Human)
  
  Details:
  HIF1 alpha antibody was used for auantitative HIF1 alpha meso scale discovery immunoassay on experiments involving RCC4, HeLa, Hep3B, MCF-7 and U2OS cell lines.
• Nakatani K, Watabe T, Masuda D et al. Myocardial energy provision is preserved by increased utilization of glucose and ketone bodies in CD36 knockout mice Metab. Clin. Exp. 2015-06-06 [PMID: 26130608] (WB, Mouse)
  
  Details:
  HIF1 alpha antibody was used for WB on protein extraction from cardiac tissues of WT and CD-36 knock-out mice (Fig. 2G).
• Lee JH, Elly C, Park Y, Liu YC. E3 Ubiquitin Ligase VHL Regulates Hypoxia-Inducible Factor-1a to Maintain Regulatory T Cell Stability and Suppressive Capacity Immunity. 2015-06-16 [PMID: 26084024] (Chemotaxis, Mouse)
• Jung J, Sosnowska D, Weaver J et al. Expression of Hypoxia-Inducible Factors in Different Stages of Pancreatic Tumor Progression Reports Oct 15 2020 (IHC, Human)
• Bohuslavova R, Skvorova L, Cerychova R, Pavlinkova G. Gene expression profiling of changes induced by maternal diabetes in the embryonic heart. Reprod Toxicol 2015-11-01 [PMID: 26073000]
• Hammami A, Charpentier T, Smans M, Stager S. IRF-5-Mediated Inflammation Limits CD8+ T Cell Expansion by Inducing HIF-1aand Impairing Dendritic Cell Functions during Leishmania Infection PLoS Pathog. 2015-06-01 [PMID: 26046638] (WB, Mouse)
• Espana-Agusti J, Tuveson DA, Adams DJ, Matakidou A. A minimally invasive, lentiviral based method for the rapid and sustained genetic manipulation of renal tubules. Sci Rep 2015-01-01 [PMID: 26046460] (IF/IHC, Mouse)
• Fan Y, Wang L, Liu C et al. Local renin-angiotensin system regulates hypoxia-induced vascular endothelial growth factor synthesis in mesenchymal stem cells. Int J Clin Exp Pathol. 2015-06-05 [PMID: 26045756] (WB, Rat)
• Sun L, Xu C, Chen G et al. A Novel Role of OS-9 in the Maintenance of Intestinal Barrier Function from Hypoxia-induced Injury via p38-dependent Pathway. Int. J. Biol. Sci. 2015-05-22 [PMID: 25999789] (WB, Human)
• Joo HY, Yun M, Jeong J et al. SIRT1 deacetylates and stabilizes hypoxia-inducible factor-1alpha (HIF-1alpha) via direct interactions during hypoxia. Biochem. Biophys. Res. Commun. 2015-05-13 [PMID: 25979359] (WB, IP, Human)
• Pang X, Wang Z, Yin D, Zhang Z. Overexpression of hypoxia-inducible factor prolyl- hydoxylase attenuated by HCG-induced vascular endothelial growth factor expression in luteal cells. Mol Med Rep 2015-05-16 [PMID: 25975603]
• Babu E, Bhutia YD, Ramachandran S et al. Deletion of the amino acid transporter Slc6a14 suppresses tumor growth in spontaneous mouse models of breast cancer. Biochem. J. 2015-05-13 [PMID: 25967569] (IHC-P, Mouse)
• Park H, Lee DS, Yim MJ. 3,3'-Diindolylmethane inhibits VEGF expression through the HIF-1alpha and NF-kB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions. Int. J. Mol. Med. 2015-05-05 [PMID: 25955241] (WB, Human)
• Takata S, Masuda T, Nakamura S et al. The effect of triamcinolone acetonide on laser-induced choroidal neovascularization in mice using a hypoxia visualization bio-imaging probe. Sci Rep. 2015-05-01 [PMID: 25927172] (WB, Mouse, Human)
• Liu X, Chen Z, Xu C et al. Repression of hypoxia-inducible factor alpha signaling by Set7-mediated methylation Nucleic Acids Res. 2015-04-20 [PMID: 25897119]
• Brandes F, Schmidt K, Wagner C, et al. Targeting cMET with INC280 impairs tumour growth and improves efficacy of gemcitabine in a pancreatic cancer model. BMC Cancer 2015-02-19 [PMID: 25884642] (WB, Human)
• Shao J, Zhang Y, Yang T et al. HIF-1alpha disturbs osteoblasts and osteoclasts coupling in bone remodeling by up-regulating OPG expression In Vitro Cell. Dev. Biol. Anim. 2015-04-10 [PMID: 25859979] (Chemotaxis, WB, Mouse)
• Chan N, He S, Spee CK et al. Attenuation of choroidal neovascularization by histone deacetylase inhibitor. PLoS One 2015-01-01 [PMID: 25807249] (WB, Human)
• Lehmann H, Vicari D, Wild PJ, Frew IJ Combined Deletion of Vhl and Kif3a Accelerates Renal Cyst Formation. J Am Soc Nephrol 2015-11-01 [PMID: 25788526]
• Wang F, Zhang G, Xing T et al. Renalase contributes to the renal protection of delayed ischaemic preconditioning via the regulation of hypoxia-inducible factor-1a. J. Cell. Mol. Med. 2015-03-17 [PMID: 25781495] (WB, Human)
• Fillmore N, Huqi A, Jaswal JS et al. Effect of Fatty acids on human bone marrow mesenchymal stem cell energy metabolism and survival. PLoS ONE. 2015-03-15 [PMID: 25768019] (WB, Human)
• Srivastava S, Thakkar B, Yeoh KG et al. Expression of proteins associated with hypoxia and Wnt pathway activation is of prognostic significance in hepatocellular carcinoma Virchows Arch. 2015-03-06 [PMID: 25742908] (IHC-P, Human)
• Paeng SH, Jung WK, Park WS et al. Caffeic acid phenethyl ester reduces the secretion of vascular endothelial growth factor through the inhibition of the ROS, PI3K and HIF-1 alpha signaling pathways in human retinal pigment epithelial cells under hypoxic conditions Int. J. Mol. Med. 2015-05-01 [PMID: 25738890] (WB, Human)
• Rees ML, Subramaniam J, Li Y et al. A PKM2 signature in the failing heart. Biochem Biophys Res Commun 2015-04-10 [PMID: 25735978]
• Hasan AU, Ohmori K, Konishi K et al. Eicosapentaenoic acid upregulates VEGF-A through both GPR120 and PPARgamma mediated pathways in 3T3-L1 adipocytes Mol. Cell. Endocrinol. 2015-02-16 [PMID: 25697344] (WB, Mouse)
• Li R, Wang Y, Yang Z et al. Hypoxia-inducible factor-1alpha regulates the expression of L-type voltage-dependent Ca(2+) channels in PC12 cells under hypoxia Cell Stress Chaperones. 2015-02-04 [PMID: 25648081] (WB, Rat)
• Beppu T, Sasaki T et al. High-uptake areas on positron emission tomography with the hypoxic radiotracer (18)F-FRP170 in glioblastomas include regions retaining proliferative activity under hypoxia. Ann Nucl Med 2015-01-05 [PMID: 25618012] (IF/IHC, Human)
• Colbert LE, Fisher SB, Balci S et al. High nuclear hypoxia-inducible factor 1 alpha expression is a predictor of distant recurrence in patients with resected pancreatic adenocarcinoma. Int J Radiat Oncol Biol Phys 2015-03-01 [PMID: 25596110] (Human)
• Sohn, E J, Won, G Et al. Upregulation of miRNA3195 and miRNA374b Mediates the Anti-Angiogenic Properties of Melatonin in Hypoxic PC-3 Prostate Cancer Cells. J Cancer 2015-01-02 [PMID: 25553085] (ICC/IF, Human)
• Wang WK, Chen MC, Leong HF et al. Connexin 43 suppresses tumor angiogenesis by down-regulation of vascular endothelial growth factor via hypoxic-induced factor-1a Int J Mol Sci. 2014-12-26 [PMID: 25548899] (ELISA, WB, Mouse)
• Hung YH, Chang SH, Huang CT et al. Inhibitor of Differentiation-1 and Hypoxia-Inducible Factor-1 Mediate Sonic Hedgehog Induction by Amyloid Beta-Peptide in Rat Cortical Neurons. Mol. Neurobiol. 2014-12-15 [PMID: 25502463] (Chemotaxis, Rat)
• Jeong S, Park H, Hong S et al. Lipophilic modification enhances anti-colitic properties of rosmarinic acid by potentiating its HIF-prolyl hydroxylases inhibitory activity. Eur. J. Pharmacol. 2014-12-04 [PMID: 25483211] (WB, Human)
  
  Details:
  HIF1 alpha antibody used for WB in experiments involving human colon carcinoma HCT116 and HT29 cells, human renal cancer UMRC2 cells and UMRC2/VHL cells (stably transfected with VHL)
• Rutz S, Kayagaki N, Phung QT et al. Deubiquitinase DUBA is a post-translational brake on interleukin-17 production in T cells. Nature. 2014-12-03 [PMID: 25470037] (WB, Mouse)
  
  Details:
  HIF1 alpha antibody used for WB on lysates of Duba+/+ and Duba-/- mouse CD4+ T(H)17 cells stimulated with anti-CD3 and anti-CD28 antibodies in the presence of TGF-b and IL-6 (Figure 7).
• Samanta D, Gilkes D, Chaturvedi P, Xiang L, Semenza G. Hypoxia-inducible factors are required for chemotherapy resistance of breast cancer stem cells. Proc Natl Acad Sci U S A. 2014-12-01 [PMID: 25453096] (WB, Human)
• Gillespie DL, Aguirre MT, Ravichandran S et al. RNA interference targeting hypoxia-inducible factor 1alpha via a novel multifunctional surfactant attenuates glioma growth in an intracranial mouse model. J. Neurosurg. 2014-11-25 [PMID: 25423275] (IHC-P, Human)
  
  Details:
  HIF1 alpha antibody used for IHC-P on sections of orthotopic mouse model with U87-LucNeo cells subjected to RNAi mediated knock down HIF-1 alpha in vivo
• Wu X, He L, Chen F et al. Impaired autophagy contributes to adverse cardiac remodeling in acute myocardial infarction. PLoS OnE. 2014-11-21 [PMID: 25409294] (WB, Mouse)
• Sato Y, Miyauchi Y, Yoshida S et al. The Vitamin D Analogue ED71 but not 1,25(OH)2D3 Targets HIF1Alpha Protein in Osteoclasts PLoS OnE et al. 2014-11-07 [PMID: 25375896] (WB, Mouse)
• Rahman Su, Lee Ms, Baek Jh et al. The Prolyl Hydroxylase Inhibitor Dimethyloxalylglycine Enhances Dentin Sialophoshoprotein Expression through VEGF-Induced Runx2 Stabilization PLoS OnE et al. 2014-11-05 [PMID: 25369078] (WB, Rat)
  
  Details:
  HIF1 alpha antibody used for WB on the whole cell lysates of MDPC-23 cells (a rat odontoblast-like cell line) incubated for 24 h in the presence or absence of DMOG 0-1 mM concentration (Figure 1B).
• Assar Me, Sanchez-Puelles Jm, Royo I et al. FM19G11 reverses endothelial dysfunction in rat and human arteries through stimulation of the PI3K/Akt/enOS pathway, independently of mTOR/HIF-1Alpha activation Br. J. Pharmacol. 2014-11-03 [PMID: 25363469] (WB, Rat)
  
  Details:
  HIF-1 alpha antibody used for WB on lysates of ex vivo FM19G11 treated aortic and mesenteric arteries from control and insulin-resistant rats (Figure 6).
• Funk K, Scheerer N, Verhaegh R et al. Severe blunt muscle trauma in rats: only marginal hypoxia in the injured area PLoS OnE et al. 2014-11-01 [PMID: 25360779] (IHC-P, Rat)
  
  Details:
  HIF1 alpha antibody used for IHC-P on right Musculus gastrocnemius tissue section of male Wistar rats subjected to severe blunt muscle trauma with standardized weight-drop device - paraformaldehyde-fixed-paraffin-embedded sections, primary antibody dilution of 1:10000, catalyzed signal amplification system from DAKO, staining development with DAB-Hematoxylin (Figure 5).
• Marhold M, Tomasich E, El-Gazzar A et al. HIF-1alpha Regulates mTOR Signaling and Viability of Prostate Cancer Stem Cells. Mol. Cancer Res. 2014-10-27 [PMID: 25349289] (WB, IP, Human, Mouse)
• He YL, Li MM, Wu LY et al. Enhanced Hypoxia-Inducible Factor (HIF)-1 alpha Stability Induced by 5-Hydroxymethyl-2-Furfural (5-HMF) Contributes to Protection against Hypoxia Mol. Med. 2015-03-27 [PMID: 25333920] (WB, ICC/IF, Rat, Mouse)
• Yoshikawa N, Shimizu N, Ojima H et al. Down-regulation of hypoxia-inducible factor-1 alpha and vascular endothelial growth factor by HEXIM1 attenuates myocardial angiogenesis in hypoxic mice. Biochem. Biophys. Res. Commun. 2014-10-06 [PMID: 25301555] (WB, Human)
• Kasper LH, Qu C, Obenauer JC et al. Genome-wide and single-cell analyses reveal a context dependent relationship between CBP recruitment and gene expression. Nucleic Acids Res. 2014-09-23 [PMID: 25249627] (Chemotaxis, Mouse)
• Bordji K, Grandval A, Cuhna-Alves L et al. Hypoxia-inducible factor-2a (HIF-2a), but not HIF-1a, is essential for hypoxic induction of class III b-tubulin expression in human glioblastoma cells. FEBS J. 2014-09-22 [PMID: 25244496] (WB, GS, Human)
  
  Details:
  HIF-2 alpha/EPAS1 antibody used for WB and EMSA/Gel Super Shift Assay in experiments involving human glioblastoma cell lines - U87MG (p53 wild-type, PTEN mutant) , U251MG (p53 mutant, PTEN mutant) and GL15 (obtained from a frontal lobe tumor biopsy of a 59-year-old man with GBM).
• Shinohara M, Kibi M, Riley IR et al. Cell-Cell Interactions and Bronchoconstrictor Eicosanoid Reduction With Inhaled Carbon Monoxide and Resolvin D1. Am. J. Physiol. Lung Cell Mol. Physiol. 2014-09-12 [PMID: 25217660] (ICC/IF, Human)
• Lee HS, Kim KS, Lim HS et al. Priming Wharton's jelly-derived mesenchymal stromal/stem cells with ROCK inhibitor improves recovery in an intracerebral hemorrhage model. J Cell. Biochem. 2014-09-04 [PMID: 25185536]
• Fujikuni N, Yamamoto H, Tanabe K et al. Hypoxia-mediated CD24 expression is correlated with gastric cancer aggressiveness by promoting cell migration and invasion. Cancer Sci 2014-11-01 [PMID: 25174257] (WB)
• Queisser MA, Dada LA, Deiss-Yehiely N. HOIL-1L Functions as the PKC-zeta Ubiquitin Ligase to Promote Lung Tumor Growth. Am. J. Respir. Crit. Care Med. 2014-08-13 [PMID: 25118570]
• Huang T, Huang W, Zhang Z et al. Hypoxia-inducible factor-1a upregulation in microglia following hypoxia protects against ischemia-induced cerebral infarction. Neuroreport. 2014-10-01 [PMID: 25089804]
• Gu J, Fan Y, Liu X et al. SENP1 protects against myocardial ischaemia/reperfusion injury via a HIF1a-dependent pathway. Cardiovasc. Res. 2014-07-31 [PMID: 25082844] (WB, IP, Rat)
• Yang Xi, Zhu Hongcheng, Ge Yangyang et al. Melittin enhances radiosensitivity of hypoxic head and neck squamous cell carcinoma by suppressing HIF-1alpha. Tumour Biol. 2014-07-23 [PMID: 25053591]
• Zhou W, Marinescu M, Veltkamp R. Only Very Early Oxygen Therapy Attenuates Posthemorrhagic Edema Formation and Blood-Brain Barrier Disruption in Murine Intracerebral Hemorrhage. Neurocrit Care 2014-07-15 [PMID: 25022804] (WB, Mouse)
  
  Details:
  HIF-1 alpha antibody used for WB (1:500 dilution) on lysates of serial coronal cryosections of hemorrhagic and nonhemorrhagic hemispheres from C57BL/6 male mice 3d after inducing Intracerebral Hemorrhage/ ICH (Fig 6B).
• Abu-Remaileh M, Aqeilan RI. Tumor suppressor WWOX regulates glucose metabolism via HIF1alpha modulation. Cell Death Differ 2014-11-01 [PMID: 25012504] (WB)
• Kalamida D, Karagounis IV, Giatromanolaki A, Koukourakis MI. Important role of autophagy in endothelial cell response to ionizing radiation. PLoS ONE 2014-07-11 [PMID: 25010689] (WB, Human)
  
  Details:
  HIF-1 alpha antibody used for WB on lysates of HUVEC/human umbilical vein endothelial cells after exposure to 2 Gy of IR with or without 2 day pre-incubation with SMER28/Small-Molecule Enhancer of Rapamycin-28 (an mTOR-independent inducer of autophagy). Primary antibody was used at 1:500 dilution and the blot images are not shown in the article.
• Privat M, Radosevic-Robin N, Aubel C et al. BRCA1 Induces Major Energetic Metabolism Reprogramming in Breast Cancer Cells. PLoS ONE 2014-07-11 [PMID: 25010005] (WB, Human)
  
  Details:
  HIF-1 alpha antibody used for WB on hypoxia mimetic CoCl2 treated /not treated SUM1315-BRCA1 and SUM1315-LXSN cell lines which were obtained by stable transfection of SUM1315 human breast cancer cells with an empty LXSN plasmid, or a BRCA1-encoding LXSN plasmid (WB image shown in Figure S1).
• Huang Miaoling, Chen Qing, Xiao Jianpeng et al. Overexpression of hypoxia-inducible factor-1alpha is a predictor of poor prognosis in cervical cancer: a clinicopathologic study and a meta-analysis. Int J Gynecol Cancer. 2014-07-01 [PMID: 24978711] (Human)
• Yang MY, Chiao MT, Lee HT et al. An innovative three-dimensional gelatin foam culture system for improved study of glioblastoma stem cell behavior J. Biomed. Mater. Res. Part B Appl. Biomater. 2014-06-26 [PMID: 24966152] (IF/IHC, Human)
  
  Details:
  Laminin antibody used for immunocytochemical/ICC staining (DAB chromogen) of human glioblastoma stem cells/GSCs grown in gelatin foam culture system - 3-D culture environment model. Figure 4 e - Laminin (cell adhesion molecule ) absent in monolayer cultures, but expressed well in GSC mass growth with gelatin cultures.
• Wu JB, Shao C, Li X et al. Near-infrared fluorescence imaging of cancer mediated by tumor hypoxia and HIF1alpha/OATPs signaling axis. Biomaterials. 2014-06-20 [PMID: 24957295] (IHC-P, Human)
  
  Details:
  Archival formalin-fixed paraffin-embedded/FFPE human prostate cancer/PCa specimens were processed for IHC staining with HIF1 alpha antibody (clone H1alpha67, 1:30 dilution).
• Narise Kosuke, Okuda Kensuke, Enomoto Yukihiro et al. Optimization of biguanide derivatives as selective antitumor agents blocking adaptive stress responses in the tumor microenvironment. Drug Des Devel Ther. 2014-06-06 [PMID: 24944508] (WB)
• Vorrink SU, Sarsour EH, Olivier AK et al. PCB 126 perturbs hypoxia-induced HIF-1a activity and glucose consumption in human HepG2 cells Exp. Toxicol. Pathol. 2014-06-07 [PMID: 24916446] (IHC-P, Rat)
  
  Details:
  HIF 1 alpha antibody used for FFPE liver sections from male Sprague-Dawley rats treated or not with PCB 126 (AhR agonist ) - HIER using citrate buffer pH 6.0, Peroxidase quenching with 3% H2O2 for 8 min, blocking with 5% horse serum, 1:50 primary - 2 h incubation at RT, detected via Mouse Envision HRP System Reagent -DAB method. Fig 1 - PCB 126 treated livers had mainly cytoplasmic/decreased nuclear staining compared to control rats.
• Salman Shaima, Holloway Alison C, Nurse Colin A. Chronic opioids regulate KATP channel subunit Kir6.2 and carbonic anhydrase I and II expression in rat adrenal chromaffin cells via HIF-2alpha and protein kinase A. Am J Physiol Cell Physiol. 2014-08-01 [PMID: 24898587] (Rat)
• Ding Z, Zhou M, Zhou Z et al. Injectable Silk Nanofiber Hydrogels for Sustained Release of Small-Molecule Drugs and Vascularization ACS Biomater. Sci. Eng. Jul 10 2019
• Zhang L, Sun Y, Fei M et al. Disruption of chaperone-mediated autophagy-dependent degradation of MEF2A by oxidative stress-induced lysosome destabilization. Autophagy 2014-06-01 [PMID: 24879151]
• Toth RK, Tran JD, Muldong MT et al. Hypoxia-Induced PIM Kinase and Laminin-Activated Integrin alpha 6 Mediate Resistance to PI3K Inhibitors in Bone-Metastatic CRPC bioRxiv (IF/IHC, Human, Mouse)
• Arreola A, Cowey CL, Coloff JL et al. HIF1alpha and HIF2alpha Exert Distinct Nutrient Preferences in Renal Cells. PLoS ONE. 2014-06-02 [PMID: 24879016] (WB, Mouse)
• Williamson DL, Li Z, Tuder RM et al. Altered nutrient response of mTORC1 as a result of changes in REDD1 expression: effect of obesity vs. REDD1 deficiency J Appl Physiol (1985). 2014-05-28 [PMID: 24876363] (WB, Mouse)
• Xiong Y, Qu Z, Chen N et al. The local corticotropin-releasing hormone receptor 2 signalling pathway partly mediates hypoxia-induced increases in lipolysis via the cAMP-protein kinase A signalling pathway in white adipose tissue. Mol. Cell. Endocrinol. 2014-05-22 [PMID: 24859650] (WB, Rat)
• Barbosa DM Effect of the rhubarb anthraquinone Rhein on cardiac fibroblast to myofibroblast transition Thesis
• Tran MGB, Bibby BAS, Yang L et al. Independence of HIF1a and androgen signaling pathways in prostate cancer bioRxiv (IF/IHC, Human)
• Xu W, Kwon JH, Moon YH et al. Influence of preoperative transcatheter arterial chemoembolization on gene expression in the HIF-1alpha pathway in patients with hepatocellular carcinoma. J. Cancer Res. Clin. Oncol. 2014-05-23 [PMID: 24853275] (IHC-P, Human)
• Schiffer Davide, Mellai Marta, Annovazzi Laura et al. Stem cell niches in glioblastoma: a neuropathological view. Biomed Res Int. 2014-04-15 [PMID: 24834433] (Human)
• Maugeri G The role of VIP family members in diabetic retinopathy Thesis (ICC/IF, IB, Rat)
• Kim YS, Lee HA, Lim JY et al. beta-Carotene inhibits neuroblastoma cell invasion and metastasis in vitro and in vivo by decreasing level of hypoxia-inducible factor-1alpha. J. Nutr. Biochem. 2014-03-13 [PMID: 24746828] (WB, IHC-P, Human)
• Bagchi R, Bagchi A, Salunke A et al. Chemical hypoxia in human pluripotent NT2 stem cell-derived neurons: Effect of hydroxamic acid and benzamide-based epigenetic drugs Neurosci Res Notes 2019-08-22 (ELISA, Human)
• Zheng J, Song F, Lu SL, Wang XQ. Dynamic Hypoxia in Scar Tissue During Human Hypertrophic Scar Progression. Dermatol Surg 2014-03-31 [PMID: 24684437] (IHC-P, Human)
• Chang ML, Chiu CJ, Shang F, Taylor A. High Glucose Activates ChREBP-Mediated HIF-1a and VEGF Expression in Human RPE Cells Under Normoxia. Adv. Exp. Med. Biol. 2014-03-25 [PMID: 24664750] (WB, Human)
• Raspaglio G, Petrillo M, Martinelli E et al. Sox9 and Hif-2a regulate TUBB3 gene expression and affect ovarian cancer aggressiveness. Gene 2014-03-21 [PMID: 24661907] (WB, Human)
• Zhang WG, He L, Shi XM et al. Regulation of transplanted mesenchymal stem cells by the lung progenitor niche in rats with chronic obstructive pulmonary disease. Respir. Res. 2014-03-25 [PMID: 24661402] (IHC-P, WB, Rat)
• Sano M, Sasaki T, Hirakawa S et al. Lymphangiogenesis and angiogenesis in abdominal aortic aneurysm. PLoS ONE 2014-03-21 [PMID: 24651519] (IHC-P, Human)
• Chen Jing, Li Tao, Liu Qilun et al. Clinical and prognostic significance of HIF-1alpha, PTEN, CD44v6, and survivin for gastric cancer: a meta-analysis. PLoS One. 2014-03-19 [PMID: 24647137]
• Pan WL, Wong JH, Fang EF et al. Preferential cytotoxicity of the type I ribosome inactivating protein alpha-momorcharin on human nasopharyngeal carcinoma cells under normoxia and hypoxia. Biochem. Pharmacol. 2014-03-14 [PMID: 24637239] (WB, Human)
• Zhao L, Yang YF, Gao YB et al. Upregulation of HIF-1a Via Activation of ERK and PI3K Pathway Mediated Protective Response to Microwave-Induced Mitochondrial Injury in Neuron-Like Cells. Mol. Neurobiol. 2014-03-14 [PMID: 24627260] (WB, Rat)
• Gauter-Fleckenstein Benjamin, Reboucas Julio S, Fleckenstein Katharina et al. Robust rat pulmonary radioprotection by a lipophilic Mn N-alkylpyridylporphyrin, MnTnHex-2-PyP(5+). Redox Biol. 2014-01-09 [PMID: 24624330] (IF/IHC, Rat)
• Shan F, Li J, Huang QY. HIF-1 Alpha-Induced Up-Regulation of miR-9 Contributes to Phenotypic Modulation in Pulmonary Artery Smooth Muscle Cells During Hypoxia. J. Cell. Physiol. 2014-02-25 [PMID: 24615545] (WB, Rat)
• Imamura Y, Tomita S, Imanishi M et al. HIF-2a/ARNT complex regulates hair development via induction of p21Waf1/Cip1 and p27Kip1. FASEB J. 2014-03-05 [PMID: 24599965] (WB, Mouse)
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  Details:
  Using the Biotin conjugated version of NB100-134, catalog number NB100-134B.
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  Details:
  Citation using the DyLight 550 version of this antibody.
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• Hayashi M, Sakata M, Takeda T et al. Up-regulation of c-met protooncogene product expression through hypoxia-inducible factor-1alpha is involved in trophoblast invasion under low-oxygen tension. Endocrinology 2005-11-01 [PMID: 16099863] (Chemotaxis, Human)
• Behan A, Doyle S, Farrell M. Adaptive responses to mitochondrial dysfunction in the rho degrees Namalwa cell. Mitochondrion 2005-06-01 [PMID: 16050983] (WB, Human)
• Zagzag D, Krishnamachary B, Yee H et al. Stromal cell-derived factor-1alpha and CXCR4 expression in hemangioblastoma and clear cell-renal cell carcinoma: von Hippel-Lindau loss-of-function induces expression of a ligand and its receptor. Cancer Res 2005-07-15 [PMID: 16024619] (WB, IF/IHC, Human)
• Maltepe E, Krampitz GW, Okazaki KM et al. Hypoxia-inducible factor-dependent histone deacetylase activity determines stem cell fate in the placenta. Development 2005-08-01 [PMID: 15987772] (WB, Mouse)
• Alberghini A, Recalcati S, Tacchini L et al. Loss of the von Hippel Lindau tumor suppressor disrupts iron homeostasis in renal carcinoma cells. J Biol Chem. 280(34):30120-8. 2005-08-26 [PMID: 15985433] (WB, Human)
• Kim SJ, Rabbani ZN, Dewhirst MW et al. Expression of HIF-1alpha, CA IX, VEGF, and MMP-9 in surgically resected non-small cell lung cancer. Lung Cancer 2005-09-01 [PMID: 15935515] (IF/IHC, Human)
• Wang, B, Zou, Y, Li, H et al. Genistein Inhibited Retinal Neovascularization Expression of Vascular Endothelial Growth Factor Hypoxia Inducible Factor 1 alpha in a Mouse Model of Oxygen-Induced Retinopathy. J. of Ocular Pharmacology Therapeutics.:107-113. 2005-04-01 [PMID: 15857276] (Mouse)
• Li YM, Zhou BP, Deng J et al. A hypoxia-independent hypoxia-inducible factor-1 activation pathway induced by phosphatidylinositol-3 kinase/Akt in HER2 overexpressing cells. Cancer Res 2005-04-15 [PMID: 15833858]
• Kazi AA, Jones JM, Koos RD. Chromatin immunoprecipitation analysis of gene expression in the rat uterus in vivo: estrogen-induced recruitment of both estrogen receptor alpha and hypoxia-inducible factor 1 to the vascular endothelial growth factor promoter. Mol Endocrinol 2005-08-01 [PMID: 15774498] (WB)
• Matsuyama T, Nakanishi K, Hayashi T et al. Expression of hypoxia-inducible factor-1alpha in esophageal squamous cell carcinoma. Cancer Sci 2005-03-01 [PMID: 15771621]
• Irie N, Matsuo T, Nagata I. Protocol of radiotherapy for glioblastoma according to the expression of HIF-1. Brain Tumor Pathol 2004-01-01 [PMID: 15696961] (IF/IHC, Human)
• Aminova LR, Chavez JC, Lee J et al. Prosurvival prodeath effects of hypoxia-inducible factor-1alpha stabilization in a murine hippocampal cell line. J Biol Chem. 280(5):3996-4003. 2005-02-04 [PMID: 15557337] (WB, Mouse)
• Knies-Bamforth, UE et al. c-Myc interacts with hypoxia to induce angiogenesis in vivo by a vascular endothelial growth factor-dependent mechanism. Can. Res. 64:6563-6570. 2004-01-01 [PMID: 15374969] (WB, Rat)
• Tezel G, Wax MB. Hypoxia-inducible factor 1alpha in the glaucomatous retina and optic nerve head. Arch Ophthalmol 2004-09-01 [PMID: 15364715] (IHC-P, Human)
• Swinson DE, Jones JL, Cox G et al. Hypoxia-inducible factor-1 alpha in non small cell lung cancer: relation to growth factor, protease and apoptosis pathways. Int J Cancer 2004-08-10 [PMID: 15185341] (IP, Human)
• Mansfield KD, Simon MC, Keith B. Hypoxic reduction in cellular glutathione levels requires mitochondrial reactive oxygen species. J Appl Physiol (1985) 2004-10-01 [PMID: 15180977] (WB, Human)
• Nishi H, Nakada T, Hokamura M et al. Hypoxia-inducible factor-1 transactivates transforming growth factor-beta3 in trophoblast. Endocrinology 2004-09-01 [PMID: 15155569] (WB, Human)
• Tissot van Patot MC, Bendrick-Peart J, Beckey VE et al. Greater vascularity, lowered HIF-1/DNA binding, and elevated GSH as markers of adaptation to in vivo chronic hypoxia. Am J Physiol Lung Cell Mol Physiol 2004-09-01 [PMID: 15132953] (WB, Human)
• Reichner JS, Albina JE. Determination of the role of hypoxia-inducible factor 1 in wound healing. Methods Enzymol 2004-01-01 [PMID: 15063696] (Rat)
• Pascual O, Roux JC, Soulage C et al. Carotid chemodenervation approach to study oxygen sensing in brain stem catecholaminergic cells. Methods Enzymol 2004-01-01 [PMID: 15063691] (IF/IHC, Rat)
• Yatabe N, Kyo S, Maida Y et al. HIF-1-mediated activation of telomerase in cervical cancer cells. Oncogene 2004-04-29 [PMID: 15048086] (Chemotaxis, Human)
• Wykoff CC, Sotiriou C, Cockman ME et al. Gene array of VHL mutation and hypoxia shows novel hypoxia-induced genes and that cyclin D1 is a VHL target gene. Br J Cancer 2004-03-22 [PMID: 15026807] (Human)
• Erler JT, Cawthorne CJ, Williams KJ et al. Hypoxia-mediated down-regulation of Bid and Bax in tumors occurs via hypoxia-inducible factor 1-dependent and-independent mechanisms and contributes to drug Mol Cell Biol. 2004-04-01 [PMID: 15024076] (WB, Mouse, Human)
• Palladino MA, Powell JD, Korah N, Hermo L. Expression and localization of hypoxia-inducible factor-1 subunits in the adult rat epididymis. Biol Reprod 2004-04-01 [PMID: 14668208] (WB, Rat)
• Hu CJ, Wang LY, Chodosh LA et al. Differential roles of hypoxia-inducible factor 1alpha (HIF-1alpha) and HIF-2alpha in hypoxic gene regulation. Mol Cell Biol 2003-12-01 [PMID: 14645546] (WB)
• Imai T, Horiuchi A et al. Hypoxia attenuates the expression of E-cadherin via up-regulation of SNAIL in ovarian carcinoma cells. Am J Pathol 2003-01-10 [PMID: 14507651] (IF/IHC, Human)
• Liu MY, Poellinger L, Walker CL. Up-regulation of hypoxia-inducible factor 2alpha in renal cell carcinoma associated with loss of Tsc-2 tumor suppressor gene. Cancer Res 2003-05-15 [PMID: 12750296] (Rat)
• Park, J et al. Gastric epithelial reactive oxygen species prevent normoxic degradation of hypoxia-inducible factor-1alpha in gastric cancer cells. Clin. Can. Res. 9:433-440. 2003-01-01 [PMID: 12538497] (WB, Human)
• Ivan, M et al. Biochemical purification and pharmacological inhibition of a mammalian prolyl hydroxylase acting on hypoxia-inducible factor. PNAS. 99: 13459-13464. 2002-01-01 [PMID: 12351678] (WB, Human)
• Des Tombe AL, Van Beek-Harmsen BJ, Lee-De Groot MB, Van Der Laarse WJ. Calibrated histochemistry applied to oxygen supply and demand in hypertrophied rat myocardium. Microsc Res Tech 2002-09-01 [PMID: 12226811] (IF/IHC, Rat)
• Powell JD, Elshtein R, Forest DJ, Palladino MA. Stimulation of hypoxia-inducible factor-1 alpha (HIF-1alpha) protein in the adult rat testis following ischemic injury occurs without an increase in HIF-1alpha messenger RNA expression. Biol Reprod 2002-09-01 [PMID: 12193413] (WB, Rat)
• Schindl M, Schoppmann SF, Samonigg H et al. Overexpression of hypoxia-inducible factor 1alpha is associated with an unfavorable prognosis in lymph node-positive breast cancer. Clin. Can. Res. 8: 1831-1837. 2002-01-01 [PMID: 12060624] (IHC-P, ICC/IF, Human)
• Ivan, M et al. Structure of an HIF-1alpha -pVHL complex: hydroxyproline recognition in signaling. Science; 292(5516): 464-8. 2001-01-01 [PMID: 12004076] (IP)
• Shoshani T, Faerman A, Mett I et al. Identification of a novel hypoxia-inducible factor 1-responsive gene, RTP801, involved in apoptosis. Mol Cell Biol 2002-04-01 [PMID: 11884613] (EMSA)
• Pascual O, Denavit-Saubie M, Dumas S et al. Selective cardiorespiratory and catecholaminergic areas express the hypoxia-inducible factor-1alpha (HIF-1alpha) under in vivo hypoxia in rat brainstem. Eur J Neurosci 2001-12-01 [PMID: 11860493] (ICC/IF, Rat)
• Arsham AM, Plas DR, Thompson CB, Simon MC. Phosphatidylinositol 3-kinase/Akt signaling is neither required for hypoxic stabilization of HIF-1 alpha nor sufficient for HIF-1-dependent target gene transcription. J Biol Chem 2002-04-26 [PMID: 11859074] (WB)
• Stokes DG, Liu G, Coimbra IB et al. Assessment of the gene expression profile of differentiated and dedifferentiated human fetal chondrocytes by microarray analysis Arthritis Rheum 2002-02-13 [PMID: 11840443] (WB, Bovine)
• Albina JE, Mastrofrancesco B, Vessella JA et al. HIF-1 expression in healing wounds: HIF-1alpha induction in primary inflammatory cells by TNF-alpha. Am J Physiol Cell Physiol 2001-12-01 [PMID: 11698256] (Rat)
• Kurebayashi J, Otsuki T, Moriya T et al. Hypoxia reduces hormone responsiveness of human breast cancer cells. Jpn J Cancer Res 2001-10-01 [PMID: 11676860] (IHC-P, Human)
• Duyndam, MCA et al. Induction of vascular endothelial growth factor expression and hypoxia-inducible factor 1alpha protein by the oxidative stressor arsenite. J Biol Chem;276(51):48066-76. 2001-12-21 [PMID: 11641398] (WB, Human)
• Schindl M, Oberhuber G, Pichlbauer EG et al. DNA repair-redox enzyme apurinic endonuclease in cervical cancer: evaluation of redox control of HIF-1alpha and prognostic significance. Int J Oncol 2001-10-01 [PMID: 11562758] (Human)
• Birner P, Gatterbauer B, Oberhuber G et al. Expression of hypoxia-inducible factor-1 alpha in oligodendrogliomas: its impact on prognosis on neoangiogenesis. Cancer;92(1):165-71. 2001-07-01 [PMID: 11443623] (IHC-P, Human)
• Birner P, Schindl M, Obermair A et al. Expression of hypoxia-inducible factor 1alpha in epithelial ovarian tumors: its impact on prognosis and on response to chemotherapy. Clin Cancer Res 2001-06-01 [PMID: 11410504] (IF/IHC, Human)
• Ivan M, Kondo K, Yang H et al. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 2001-04-20 [PMID: 11292862]
• Jaakkola P, Mole DR, Tian YM et al. Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation. Science 2001-04-20 [PMID: 11292861] (Human)
• Birner P, Schindl M, Obermair A et al. Overexpression of hypoxia-inducible factor 1alpha is a marker for an unfavorable prognosis in early-stage invasive cervical cancer. Cancer Res;60(17):4693-6. 2000-09-01 [PMID: 10987269] (IHC-P, Human)
• Kamura, T et al. Activation of HIF1alpha ubiquitination by a reconstituted von Hippel-Lindau (VHL) tumor suppressor complex. PNAS 97: 10430-10435. 2000-01-01 [PMID: 10973499] (WB, IP)
• Wang G, Hazra TK, Mitra S et al. Mitochondrial DNA damage and a hypoxic response are induced by CoCl(2) in rat neuronal PC12 cells. Nucleic Acids Res 2000-05-15 [PMID: 10773083] (WB)
• Sutter, CH et al. Hypoxia-inducible factor 1alpha protein expression is controlled by oxygen-regulated ubiquitination that is disrupted by deletions and missense mutations. PNAS. 97: 4748-4753. 2000-01-01 [PMID: 10758161] (WB, Human)
• Gothie, E et al. Identification of alternative spliced variants of human hypoxia-inducible factor-1alpha. J. Biol. Chem. 275(10):6922-6927. 2000-01-01 [PMID: 10702253] (WB, Human)
• Zhong, H et al. Overexpression of hypoxia-inducible factor 1alpha in common human cancers and their metastases. Can. Res.1999; 59: 5830-5835. 1999-01-01 [PMID: 10582706] (IHC-P, Mouse, Human)
• Palladino MA, Powell JD, Korah N et al. Expresion Localization of Hypoxia-Inducible Factor-1 Subunits in the Adult Rat Epididymis. Biol Reprod. 70(4):1121-30. 2004-04-01 [PMID: 1466820]

Reviews for HIF-1 alpha Antibody (NB100-105) (44)

Average Rating: 4.2

(Based on 44 reviews)

We have 44 reviews tested in 5 species: Human, Mouse, Rat, Other, 0.

reviewed by: Verified Customer IHC-P Human 01/19/2022

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Throat cancer tissue
• Species: Human
• Lot: H1alpha67

Comments

• Comments: PFA fixed paraffin-embedded sections. Primary antibody dilution 1:100.

reviewed by: Verified Customer WB Human 03/31/2021

Summary

• Application: Western Blot
• Sample Tested: Caki-1 cell lysate,fibroblasts
• Species: Human
• Lot: BQ

reviewed by: Verified Customer WB Human 08/09/2020

Summary

• Application: Western Blot
• Sample Tested: HEK293A whole cell lysate,RCC4 Whole cell lysate
• Species: Human
• Lot: BH-23

reviewed by: Verified Customer WB Human 06/13/2019

Summary

• Application: Western Blot
• Sample Tested: Blood mononuclear cells (PBMCs)
• Species: Human
• Lot: BH-13

Comments

• Comments: Confirmation of induction of hypoxia in CoCl2 treated PBMCs infected with vGPCR frame shift knock out and wild type revertant KSHV virion. KSHV infected cells were treated with CoCl2 for 48 hours followed by protein isolation and western blot against HIF1 alpha .

reviewed by: Verified Customer WB Human 02/02/2019

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell whole cell lysate
• Species: Human
• Lot: AB6

reviewed by: Verified Customer WB Rat 02/02/2019

Summary

• Application: Western Blot
• Sample Tested: Rat liver
• Species: Rat
• Lot: AW1

reviewed by: Verified Customer WB Human 01/29/2019

Summary

• Application: Western Blot
• Sample Tested: human adrenal cancer cells (whole cell lysate)
• Species: Human
• Lot: unknown

reviewed by: Verified Customer WB Rat 01/29/2019

Summary

• Application: Western Blot
• Sample Tested: Rat liver
• Species: Rat
• Lot: AW-1

Comments

• Comments: Detection Method: ECL

reviewed by: Verified Customer WB Human 01/24/2019

Summary

• Application: Western Blot
• Sample Tested: Hela whole cell lysate
• Species: Human
• Lot: AB7

Comments

• Comments: ECL solution for 5min, then exposure to film 3-5min

reviewed by: Verified Customer WB Human 01/23/2019

Summary

• Application: Western Blot
• Sample Tested: 293t cell lysate
• Species: Human
• Lot: NB100-105

reviewed by: Verified Customer WB Human 06/29/2017

Summary

• Application: Western Blot
• Sample Tested: 293t cell lysate
• Species: Human
• Lot: bh-1

reviewed by: Verified Customer WB Mouse 02/01/2017

Summary

• Application: Western Blot
• Sample Tested: CD4 T cells cultured in vitro, stimulated with anti-CD3/anti-CD28
• Species: Mouse

Comments

• Comments: Figure Legend: Naïve CD4 T cells from WT, VHL-deficient (Vhl KO), or HIF-1a–deficient (Hif1a KO) mice were differentiated under IL-22–skewing conditions for a total of 60 h. Some cells remained at normoxia for the duration of the culture (N); others were at normoxia for 35 h and then hypoxia (1% O2) for 24 h (H). At 60 h, nuclear extracts were harvested, and HIF-1a and lamin B1 levels were analyzed by Western blot.

reviewed by: michael dodd WB Mouse 08/17/2016

Summary

• Application: Western Blot
• Sample Tested: HL-1 cardiomyocytes
• Species: Mouse

reviewed by: Verified Customer WB Human 05/08/2016

Summary

• Application: Western Blot
• Sample Tested: k562 cells
• Species: Human
• Lot: BA-3

reviewed by: Verified Customer WB Human 04/18/2016

Summary

• Application: Western Blot
• Sample Tested: HEK293A whole cell lysate
• Species: Human
• Lot: AW-1

reviewed by: MENG xing WB Mouse 03/24/2016

Summary

• Application: Western Blot
• Species: Mouse
• Lot: BD-10

reviewed by: Verified Customer WB Mouse 10/07/2015

Summary

• Application: Western Blot
• Sample Tested: Cell lysate
• Species: Mouse

reviewed by: Zhongxiao Wang WB Mouse 10/05/2015

Summary

• Application: Western Blot
• Sample Tested: mouse retina
• Species: Mouse
• Lot: NB100-479

reviewed by: Verified Customer WB Human 08/23/2015

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell whole cell lysate
• Species: Human

reviewed by: Verified Customer WB Human 08/10/2015

Summary

• Application: Western Blot
• Sample Tested: Human prostate cancer cell line protein samples
• Species: Human

reviewed by: Verified Customer WB Human 04/28/2015

Summary

• Application: Western Blot
• Species: Human

reviewed by: Verified Customer WB Mouse 12/17/2014

Summary

• Application: Western Blot
• Sample Tested: See PIMD: 23148210
• Species: Mouse

reviewed by: Xiuquan Luo WB Human 07/18/2014

Summary

• Application: Western Blot
• Sample Tested: Caki-1 cell lysate
• Species: Human

reviewed by: Verified Customer WB Human 07/01/2014

Summary

• Application: Western Blot
• Sample Tested: Hela Cell Lysate
• Species: Human

reviewed by: Jessica Marinello WB Human 05/09/2014

Summary

• Application: Western Blot
• Sample Tested: Hela whole cell lysate
• Species: Human

reviewed by: Verified Customer WB Mouse 04/28/2014

Summary

• Application: Western Blot
• Sample Tested: mouse brain lysate
• Species: Mouse

reviewed by: Lindsy Dickerson WB Mouse 04/28/2014

Summary

• Application: Western Blot
• Sample Tested: Mouse
• Species: Mouse

reviewed by: Ewa Kaniewska IHC Other 04/14/2014

Summary

• Application: Immunohistochemistry
• Species: Other

reviewed by: Ewa Kaniewska IF Other 04/14/2014

Summary

• Application: Immunofluorescence
• Species: Other

reviewed by: Darlah Lopez WB Mouse 03/28/2014

Summary

• Application: Western Blot
• Species: Mouse

reviewed by: Hua Li WB Human 08/23/2012

Summary

• Application: Western Blot
• Sample Tested: Whole cell lysate
• Species: Human
• Lot: AB7

reviewed by: Verified Customer IF Rat 08/22/2012

Summary

• Application: Immunofluorescence
• Sample Tested: RAT KIDNEY
• Species: Rat
• Lot: F2

reviewed by: Camilla Sampaoli WB Human 08/22/2012

Summary

• Application: Western Blot
• Sample Tested: human adrenal cancer cells (whole cell lysate)
• Species: Human

reviewed by: Haofeng Ji WB Mouse 05/15/2012

Summary

• Application: Western Blot
• Sample Tested: Cell lysate
• Species: Mouse

reviewed by: Claudia Di Serio WB Human 05/11/2012

Summary

• Application: Western Blot
• Sample Tested: Human prostate cancer cells
• Species: Human
• Lot: AB1

reviewed by: Verified Customer WB Rat 12/14/2011

Summary

• Application: Western Blot
• Sample Tested: Rat liver
• Species: Rat

reviewed by: Verified Customer WB Mouse 12/09/2011

Summary

• Application: Western Blot
• Sample Tested: Mouse auditory cultured cell, Sample Amount: 30ug
• Species: Mouse

reviewed by: Verified Customer WB Human 11/21/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

reviewed by: Lu-Hai Wang ICC Human 11/15/2011

Summary

• Application: Immunocytochemistry
• Sample Tested: Human tissues
• Species: Human

reviewed by: Verified Customer WB Human 09/30/2011

Summary

• Application: Western Blot
• Sample Tested: MCF-7, Sample Amount: 100ug
• Species: Human

reviewed by: Verified Customer WB Rat 09/01/2010

Summary

• Application: Western Blot
• Sample Tested: endothelial cell nuclear extract, Sample Amount: 40ug
• Species: Rat

reviewed by: William Lin WB Mouse 05/17/2010

Summary

• Application: Western Blot
• Sample Tested: mouse cortical neurons, Sample Amount: 40ug
• Species: Mouse
• Lot: AA1

reviewed by: amy chamberlain WB 25037 04/13/2010

Summary

• Application: Western Blot
• Sample Tested: HeLa cells
• Species: 25037
• Lot: N/A

reviewed by: Verified Customer ICC Other 02/10/2010

Summary

• Application: Immunocytochemistry
• Sample Tested: HUVEC & Candida albicans
• Species: Other
• Lot: Z5

FAQs for HIF-1 alpha Antibody (NB100-105). (Showing 1 - 10 of 30 FAQs).

1. Why is there a difference between the theoretical MW for HIF1A and the observed MW for HIF-1 alpha?
   • HIF1A, like many other proteins, has post-translational modifications. Depending on the size, amount and nature of the post-translational modifications, it can cause subtle to very large changes in molecular weight.
2. Which antibody(ies) do you recommend for the detection of HIF-1a by immunohistochemistry in the sections of paraffin-embedded mouse liver samples? I would appreciate if you can give me several choices and rank them in the order of performance. My goal is to distinguish HIF upregulation by prolyl hydroxylase inhibitor in different liver cells.
   • All of our antibodies are of high quality and are well tested/validated in species/applications we list on the datasheet. However, we suggest the following four HIF-1 alpha antibodies based upon customer reviews, as well as the number of peer reviewed publications in which these products have been cited by researchers from reputed institutes. (1) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-105) (cited in at least 218 peer reviewed publications) (2) HIF-1 alpha Antibody (cat# NB100-479) (cited in at least 51 peer reviewed publications) (3) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-123 ) (cited in at least 38 peer reviewed publications) (4) HIF-1 alpha Antibody (cat# NB100-449) (cited in at least 31 peer reviewed publications).
3. I would like to know, does a path exist for detection of HIF 1 in venous blood before and after revascularization of the leg? 
   • We are not entirely sure if HIF-1 alpha will be present in the leg after revascularization. It may be present, but you may want to search the literature to see if this has been looked at before. If not, then this would certainly be an experiment worth doing.
4. What is the molecular weight (kDa) of protein HIF 1 alpha in western blot?
   • The theoretical molecular weight of HIF 1-alpha is ~93kDa. However, you will likely see a band between 100-120kDa due to phosphorylation.
5. We got the Hif1a (NB100-105) antibody from you guys. I used the concentration that is mentioned on your website, but I am getting a band of a completely different size (~70kDa) and not the 120 kDa mentioned.
   • HIF-1 alpha is a notoriously difficult protein to work with due to its rapid degradation. Therefore, the ~70kDa bands are most likely degradation products. It is very important to lyse the cells in hypoxic conditions. We strongly recommend lysing the cells directly into the Laemmli buffer and doing that quickly, so that the exposure to oxygen is minimized.Please go through our hypoxia related FAQs, you should find them very informative.Also, running a positive control may help confirm the band specificity in your samples. You may prepare them yourself or choose some from our catalog, for example: 1) HeLa Hypoxic / Normoxic Cell Lysate (NBP2-36452)2) HeLa Hypoxic (CoCl2) / Normoxic Cell Lysate (NBP2-36450)
6. I performed several Western Blots of HIF-1 alpha with different lysis buffers, whole lysates, and cytoplasm/nuclei extractions. I can’t seem to get a good western blot (poor signal, band much lower than expected, etc.). Can someone suggest some technical considerations/tricks I should consider using?
   • A major issue that researchers working with HIF-1 alpha is degradation due to exposure to oxygen. In western blot, this results in a weaker band and/or the appearance of multiple low molecular weight bands (40-80 kDa). We recommend preparing the lysates after collection of cells/tissues as quickly as possible (on ice), preferably in a hypoxic chamber. We also recommend including a true hypoxia mimetic control (eg: cells treated with CoCl2, DMOG… etc.). The controls help distinguish your band of interest from potential degradation/dimer bands.For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs.
7. I am doing HIF1 westerns in HIF-overexpressing mouse liver and adipose tissue using Novus antirabbit HIF1a antibody with overnight incubation. I am getting strong bands around 90kDa. I am aware that HIF theoretical molecular weight is 93kDa, but in westerns, the HIF band is usually around 120kDa according to my internet research. Can someone let me know if I’m getting the right HIF band or just some non-specific bands? Thanks.
   • (1)    HIF-1 alpha’s theoretical molecular weight is 93kDa. The post translationally modified/ubiquitinated form of HIF-1 alpha protein (fails to undergo proteasomal degradation) shows up as a band in the 110-130 kDa range on a Western blot.(2)    The dimeric protein may appear at a position above 200 kDa on non-reducing gels.(3)    Importantly, HIFs are among the most rapidly degradable proteins; therefore, sample preparation is highly important when analyzing HIF1 alpha or HIF2 alpha. When degraded, HIF-1 alpha may show up between 40-80 kDa position on Western blot. Degradation may be avoided by preparing the samples as soon as possible after collection of cells/tissues in hypoxic chamber. Notably, the tissues/cells should be kept on ice during lysate preparation and the lysates should be analyzed as soon as possible.(4)    For troubleshooting suggestions/feedback on more than 25 similar frequently asked questions, I would recommend visiting Novus page: FAQs - Hypoxia and HIFs (5)    Last but not the least, Novus technical support team may be contacted via email
8. I have Hif1a nuclear protein extract at -80C. I am wondering if anyone knows how long it would be good for at that temperature since HIf1a is known to be degraded easily.Thank you!
   • You could try a few things to further inhibit the degradation.1) Use the protease inhibitors (if you are not already using them).2) Lyse cells into a buffer that contains SDS or LDS (eg: Laemmli's buffer), since SDS and LDS denature and inhibit proteases. Lysis may even be performed with reducing agents in the buffer (eg. DTT), but this will make your lysates unsuitable for BCA assay.3) Lysing samples rapidly ensures that the samples are instantly homogenized (it also shears DNA released by the SDS).5) Flash-freezing samples in liquid nitrogen rather than freezing at -80*C reduces the window of time for protease activity.6) Freeze samples in individual aliquots, instead of thawing the same vial multiple times.
9. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
   • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs.
10. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
    • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs.
11. Is cross-reactivity with HIF-2 alpha tested/predicted?
    • Although we don’t have cross-reactivity data with regards to HIF-2 alpha, we predict minimal cross-reactivity based on low sequence similarity observed from BLAST analysis between HIF-1 alpha and HIF-2 alpha.
12. Do you have any suggested IHC Protocol for this antibody? We had been trying to validate protocols from other research studies but failed to get good results. Background staining seems to be the problem. 
    • Yes, we have a specific IHC protocol for NB100-105. It is located on the datasheet under the "Protocols & FAQs" tab. We also have over 300 publications that utilize this product. If you need help finding a specific paper or if you would like us to help troubleshoot your experiment, please contact Technical Support.
13. Has anyone used this antibody for supershft in EMSA?
    • Yes, this antibody is useful for EMSA and has been documented in couple of recent research publications. You may refer to the following paper for details: Kong D, et al. Echinomycin, a small-molecule inhibitor of hypoxia-inducible factor-1 DNA-binding activity. Cancer Res. 2005 Oct 1;65(19):9047-55 [PMID: 16204079].
14. What is the difference in the following two HIF-1 alpha antibodies? Catalog # NB100-105 and NB100-134
    • NB100-105 is a mouse monoclonal antibody, whereas NB100-134 is a rabbit polyclonal. They both use the same immunogen.
15. I am working on a WB using the antibody against mouse HIF1a and HIF2a (catalog numbers NB100-105 and NB100-122). I tried several anti-mouse and anti-rabbit HRP secondary antibodies (from another company) and found they gave different patterns. Please provide the secondary antibody info which was used in the WB showed on the datasheet.
    • We use catalog numbers NB7570 and NB7183 as our secondary antibodies for testing these antibodies in WB.
16. What is the difference between NB100-105 and NB100-123?
    • NB100-105 and NB100-123 are similar as far as immunogens and clones are concerned. However, they are different with respect to their purification process, concentrations and recommended dilutions. NB100-123 is Protein A purified IgG2b with a concentration of 1.5 mg/ml, whereas NB100-105 is Protein G purified IgG2b with 4.0 mg/ml concentration. Another difference is the recommended dilutions (because of the differences in their concentration).
17. Can I get a sample of the HIF1A antibody (NB100-105)?
    • We do offer this antibody in a sample size which contains 0.025ml of antibody. The sample size catalog number is NB100-105SS.
18. I did staining of 150uM Cobalt chloride induced 3T12 for 24 hours using this HIF1alpha antibody. I used a 1:50 dilution for one hour at room temperature. Should I do an overnight incubation?
    • We do recommend incubating samples overnight at 4 degrees Celsius for the best results.
19. Is there a product available to be used as the blocking peptide of NB100-105?
    • We unfortunately do not have a blocking peptide for NB100-105. It was raised against a fusion protein. We also unfortunately do not sell the protein.
20. Could you advise if I can use NBP1-96969 Ab as a control Ab for HIF1a for ChIP assay. I have a problem that normal rabbit IgG that usually are provided in the ChIP kits show higher binding compare to NB100-105 HIF1A Ab. Could you suggest if it is better to use Mouse IgG 2b Isotype control in this case or normal rabbit IgG should work as well? NBP1-96969 Ab are not validated for ChIP, please, let me know if there is something better? 
    • Yes, it would be reasonable to use a mouse IgG2b control for NB100-105 (HIF1 alpha) in ChIP assay. The control provided in the ChIP kit is recommended as a control for the histone antibodies (which are rabbit polyclonal). NBP1-96969 would be a good choice for control for the mouse monoclonal NB100-105. It has not yet been validated for ChIP, but it has been validated for use in IP, so I think it will be a good choice for control.
21. Since HIF-1 is nuclear protein, why is there obvious staining in the cytoplasm, which is displayed in the IHC image on the product website. Can you be so kind to help me? 
    • HIF1-alpha is found at very low levels in the cytoplasm under normoxic conditions. During hypoxia, HIF1-alpha is stabilized and translocates to the nucleus to act as a transcription factor. You would therefore expect to see nuclear staining in your hypoxic samples, but may also observe faint cytoplasmic staining in relation to degraded HIF in the samples. HIF proteins are notoriously difficult to work with, as they degrade very rapidly. The cytoplasmic staining is a result of HIF1-alpha degradation. I hope that this helps. You may find our guide to working with hypoxia antibodies to be useful.
22. I would like to know why I have to dilute 1:500 the primary antibody. How can I know which is the best dilution (or what amount of antibody I need to take) to my sample in the WB?
    • Our recommended dilution for use of NB100-105 in western blot is 1:500. You may need to adjust this dilution depending on the signal you are able to detect in your samples. I would recommend using a titer with 1:500 as the starting point to find the optimal dilution to use in your experiment.
23. I am wondering if you sell a blocking peptide for the HIF1a antibody cat # NB100-105?
    • We unfortunately do not sell the blocking immunogen for our product NB100-105. I am sorry for any inconvenience.
24. Could you tell me the epitope of #NB100-105?
    • We seldom map our antibodies, including monoclonals. Therefore, we do not know where this monoclonal antibody binds to in HIF-1 alpha. However, the region containing the epitope for NB100-105 could be located in the a.a. sequence used as the immunogen for raising the antibody. According to the datasheet, the immunogen for NB100-105 was: "... amino acids 432-528 of human HIF-1 alpha. [UniProt# Q16665]."
25. Would you be able to share the concentration of NB100-105 – HIF-1 alpha mAb from Lot# AB1?
    • The concentration of NB100-105 Lot AB1 is 4.0mg/ml.
26. Could you tell us if these antibodies need to use cell lysate treated with special extraction for WB? Do you have the recommended preparation method of cell lysate?
    • Usually HIF1 alpha and HIF2 alpha are hard to detect endogenously and require some inducement. The most common is hypoxic treatment of the cells. This can be also be chemically induced by CoCl2 treatment as well. Our recommended protocols and lysate preparations can be found in our white paper here
27. Is this antibody used for intracellular staining?
    • NB100-105 detects HIF-1 alpha, which is a nuclear protein, so you would want to consult our intracellular nuclear target flow protocol
28. I am using HIF1 alpha antibody NB100-105 for Western blot. Do you have any recommendations on the preparation of lysates? Also, what positive control do you suggest for WB with HIF1 alpha antibody NB100-105?
    • HIF1a protein is known to degrade easily in presence of oxygen.  For this reason the conditions for lysate preparation are crucial, we strongly recommend lysing cells quickly under cold condition by putting cell plates on ice and saving the lysates directly into the SDS-loading buffer. If nuclear fraction is desired we recommend using hypotonic buffer. You can see specific instructions for WB with HIF1 alpha antibody NB100-105 here. We recommend lysate from cells cultured in hypoxic conditions or in presence of CoCL2, which stabilizes HIF1a. We also have several controls available on our website: HeLa Hypoxic/Normoxic Cell Lysate: NBP2-36452; HeLa Hypoxic (CoCl2)/Normoxic Lysate: NBP2-36450; etc.
29. We see high background in our western blots using GAPDH antibodies H00002483-M01 and NB100-105.
    • To avoid multiple bands, we suggest:1)     Using a nitrocellulose membrane. It produces less background compared to PVDF membranes.2)     Overnight blocking at 4℃.3)     Increasing the wash time. Try washing the membrane for 10 mins, repeating 4 times.
30. NB100-105 has the same clone name (H1alpha67) as NB100-123.Are these the same clone?
    • These are the same clones, however their purifications are slightly different (and thus their recommended dilutions for certain applications are a bit different, please see the section "Applications/Dilutions" on the specific product pages for differences).NB100-123: Protein A purifiedThe basis for purification of IgG, IgG fragments and IgG subclasses is the high affinity of protein A and protein G for the Fc region of polyclonal and monoclonal IgG-type antibodies. Protein A and protein G are bacterial proteins, which, when coupled to chromatography matrix such as Agarose or Sepharose, generate exceptionally useful, easy to use media (resin) for many routine applications. Protein A/G is a recombinant of Protein A and Protein G that has the additive binding properties of both proteins. Protein A, protein G and protein A/G can be used for purification of monoclonal IgG-type antibodies, purification of polyclonal IgG subclasses, and the adsorption and purification of immune complexes involving IgG. IgG subclasses can be isolated from cell culture supernatants and serum and from ascites fluid.Lastly, we have been producing NB100-105 for a longer time period, and thus why it has considerably more publication references than NB100-123.
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Bioinformatics

• Gene Symbol: HIF1A
• Entrez:
  • 3091(Human)
• OMIM:
  • 603348(Human)