OCT4 Antibody - BSA Free

Western Blot: OCT4 Antibody [NB100-2379] - YM155 significantly reduced tumorsphere formation and inhibited EGFR autophosphorylation and G9a expression. YM155 is reported to suppress cancer stemness; therefore, we used this compound to investigate the cellular mechanism of tumorsphere formation. To compare the cytotoxic capacity of YM155 against tumorsphere formation and parental cell lines, YM155 was applied to HCC827 and A549 cells in the stemness cultured or integrated medium. (F) Moreover, YM155 inhibited EGFR autophosphorylation in A549-derived tumorspheres and reduced Oct4 expression. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0182149) licensed under a CC-BY license.

Western Blot: OCT4 Antibody [NB100-2379] - WB detection of OCT4 in mouse brain lysate using OCT4 antibody with ECL based detection. The antibody generated a specific band of OCT4 at the expected ~40kDa position.

Flow Cytometry: OCT4 Antibody [NB100-2379] - An intracellular stain was performed on Jurkat with NB100-2379 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody.

Immunocytochemistry/Immunofluorescence: OCT4 Antibody [NB100-2379] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- NB100-2379 at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Western Blot: OCT4 Antibody [NB100-2379] - MDA-MB-231 subclones transfected with NTC or HIF-1 alpha shRNA vector were treated with vehicle (V) or paclitaxel (P) and Western blot assay was performed. WB image submitted by a verified customer review.

Immunohistochemistry-Paraffin: OCT4 Antibody [NB100-2379] - Analysis of a FFPE tissue section of mouse brain using 1:200 dilution of OCT4 antibody. The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.

Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379] - An intracellular stain was performed on U2-OS cells with OCT4 NB100-2379AF647 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Western Blot: OCT4 Antibody [NB100-2379] - The expression of pluripotency factors Oct4 in human head and neck cancer cell lines.

Flow Cytometry: OCT4 Antibody [NB100-2379] - An intracellular stain was performed on Jurkat cells with OCT4 Antibody NB100-2379 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Flow Cytometry: OCT4 Antibody [NB100-2379] - An intracellular stain was performed on Neuro2a cells with OCT4 Antibody NB100-2379 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Flow Cytometry: OCT4 Antibody [NB100-2379] - An intracellular stain was performed on Ntera2 cells with OCT4 Antibody NB100-2379 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

OCT4 was detected in immersion fixed U-2 OS human osteosarcoma cell line using Rabbit anti-OCT4 Affinity Purified Polyclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB100-2379AF488) (green) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

An intracellular stain was performed on A431 human skin carcinoma cell line using Rabbit anti-OCT4 Affinity Purified Polyclonal Antibody conjugated to DyLight 650 (Catalog # NB100-2379C, blue histogram) or matched control antibody (orange histogram) at 2.5 µg/mL for 30 minutes at RT.

Western Blot: OCT4 Antibody - BSA Free [NB100-2379] - Protein expression of NANOG a, OCT4 b & SOX2 c in bovine uterine tissues obtained from control cows & from cows with adenomyosis. Data were normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Bars represent the mean ± SEM. There were no statistical differences between uterine normal & adenomyotic tissues (P > 0.05), as determined by Student’s t-test. Representative blots for NANOG, OCT4, SOX2 & GAPDH are shown below the graphs d. MM – molecular weight marker, C – tissues obtained from control cows, ADENO – tissues obtained from cows with adenomyosis Image collected & cropped by CiteAb from the following publication (//www.rbej.com/content/13/1/110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: OCT4 Antibody - BSA Free [NB100-2379] - Protein expression of pluripotency markers in uterine cells isolated from control cows & from cows with adenomyosis. NANOG a OCT4 b & SOX2 c protein expression in uterine stromal cells. NANOG d OCT4 e & SOX2 f protein expression in uterine myometrial cells. Data were normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Bars represent the mean ± SEM. Asterisks indicate statistical differences between uterine normal & adenomyotic tissue (*P < 0.05; **P < 0.01), as determined by Student’s t-test. Representative blots for NANOG, OCT4, SOX2 & GAPDH are shown below the graphs g. MM – molecular weight marker, C – cells obtained from control cows, ADENO – cells obtained from cows with adenomyosis Image collected & cropped by CiteAb from the following publication (//www.rbej.com/content/13/1/110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

An intracellular stain was performed on U-2 OS human osteosarcoma cell line with Rabbit anti-OCT4 Affinity-purified Polyclonal Antibody conjugated to Phycoerythrin (Catalog # NB100-2379PE, blue histogram) or matched control antibody (Catalog # NBP2-24983, orange histogram) at 2.5 µg/mL for 30 minutes at RT.

• Reactivity: Hu, Mu, Rt, Bv, Pm, Po, Fe
• Applications: WB, Simple Western, Flow, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

OCT4 Antibody - BSA Free Summary

• Immunogen: A synthetic peptide made to an internal portion of the human OCT4 protein (between residues 100-200) [UniProt# Q01860]
• Localization: Nuclear
• Marker: Embryonic Stem Cell Marker
• Predicted Species: Porcine (93%), Feline (93%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: POU5F1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:50-1:200
  • Immunocytochemistry/ Immunofluorescence 1:50-1:200
  • Immunohistochemistry 1:200. Use reported in scientific literature (PMID 30726734)
  • Immunohistochemistry-Paraffin 1:200
  • Simple Western
  • Western Blot reported by customer review
• Application Notes: In Western blot, a band is seen at ~40 kDa.
• Theoretical MW: 40 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:

  OCT4 Overexpression Lysate
  HeLa Whole Cell Lysate

• Control Peptide:

  OCT4 Peptide (NB100-2379PEP)

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: Feline (93%), Porcine (93%). Customer feedback suggests that this antibody does not give good results in IHC-P on feline tissues. Bovine reactivity reported in scientific literature (PMID: 23054358).

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for OCT4 Antibody - BSA Free

• class 5, transcription factor 1
• MGC22487
• Oct-3
• OCT3Oct4
• Oct-4
• Octamer-binding protein 3
• otc-4
• OTF3POU domain class 5, transcription factor 1
• OTF4
• POU class 5 homeobox 1
• POU-type homeodomain-containing DNA-binding protein

Background

OCT4 (octamer-binding protein 4) is a transcription factor that binds to the octamer motif (5'-ATTTGCAT-3') and generates a trimeric complex with SOX2 on DNA for controlling the expression of several genes (YES1, FGF4, UTF1, ZFP206 etc.) involved in embryonic development. Expressed mainly in the nucleus, OCT4 interacts with UBE2I, ZSCAN10, PKM, WWP2 etc. and OCT4-PKM interaction positively regulate OCT4's transcriptional activity. Once translated, its modifications such as sumoylation enhances the protein stability, DNA binding and transactivation activity and is required for enhanced YES1 expression. OCT4 undergoes 'Lys-63'-linked polyubiquitination by WWP2 leading to proteasomal degradation. OCT4 is found in oocytes and is zygotically expressed in pluripotent cells of pre-gastrulation embryos, and has been observed to be essential for early embryogenesis as well as for embryonic stem cell pluripotency. OCT4 is expressed in developing brain with highest levels in specific cell layers of the cortex, the olfactory bulb, hippocampus and cerebellum. Its expression decreases gradually after embryoid body formation, however, improper re-activation of OCT4 contributes to oncogenic processes and it has been implicated in certain malignancies such as breast and ovarian cancer.

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 OCT4 Antibody (NB100-2379)(30)

Publications using NB100-2379

• Xi, S;Xue, B;Wang, Y;Shan, Z;Song, T;Sun, J;Qin, S; Generation of patient-derived induced pluripotent stem cell line UJSi004-A from ultra-treatment-resistant schizophrenia Stem cell research 2024-10-09 [PMID: 39423675]
• Wei Q, Liu Z, Zhu J et Al. The Ubiquitin E3 Ligase FBXO33 Suppresses Stem Cell-Like Properties and Metastasis in Non-Small-Cell Lung Cancer by Promoting Ubiquitination and Degradation of Myc Front Biosci (Landmark Ed) 2024-08-29 [PMID: 39206900]
• Sun, T;Chen, B;Zhang, D;Xu, X;Gao, J;Sun, J;Wei, M; Generation of hereditary spastic paraplegia patient-derived induced pluripotent stem cell line UJSi003-A Stem cell research 2024-08-13 [PMID: 39154415]
• Tang S, Jones C, Dye J, Coward K Dissociation, enrichment, and the in vitro formation of gonocyte colonies from cryopreserved neonatal bovine testicular tissues Theriogenology 2023-07-22 [PMID: 37499372] (ICC/IF, Bovine)
• Zhang R, Liu L, Wang F et al. AKAP8L enhances the stemness and chemoresistance of gastric cancer cells by stabilizing SCD1 mRNA Research Square 2022-08-25 [PMID: 36522343] (WB, Human)
• Yang Y, Chen C, Zuo Q Et al. NARF is a hypoxia-induced coactivator for OCT4-mediated breast cancer stem cell specification Sci Adv 2022-12-09 [PMID: 36490339] (WB, Human)
  
  Details:
  Citation using the DyLight 405 version of this antibody.
• Panda DK, Bai X, Zhang Y et al. SCF-SKP2 E3 ubiquitin ligase links mTORC1-ER stress-ISR with YAP activation in murine renal cystogenesis The Journal of clinical investigation 2022-11-03 [PMID: 36326820] (WB, Mouse)
• Zhang R, Zhang X, Zhang W et al. Sohlh2 regulates the stemness and differentiation of colon cancer stem cells by downregulating LncRNA-H19 transcription Molecular cancer research : MCR 2022-10-26 [PMID: 36287177] (WB, Human)
• Hekman KE, Koss KM, Ivancic DZ et al. Autophagy Enhances Longevity of Induced Pluripotent Stem Cell-Derived Endothelium via mTOR-Independent ULK1 Kinase Stem cells translational medicine 2022-09-29 [PMID: 36173887] (FLOW, Human)
• Sun J, Zeng R, Liu Y, Xi S et Al. Generation of ultra-treatment-resistant schizophrenia patient-derived induced pluripotent stem cell line UJSi002-A Stem Cell Res 2022-04-03 [PMID: 35367693]
• Haiquan Lu, Yajing Lyu, Linh Tran, Jie Lan, Yangyiran Xie, Yongkang Yang, Naveena L Murugan, Yueyang J Wang, Gregg L Semenza HIF-1 recruits NANOG as a coactivator for TERT gene transcription in hypoxic breast cancer stem cells. Cell reports 2022-02-10 [PMID: 34592152]
• BAAth M, JOnsson JM, Westbom Fremer S et al. MET Expression and Cancer Stem Cell Networks Impact Outcome in High-Grade Serous Ovarian Cancer Genes 2021-05-14 [PMID: 34069138] (WB, Human)
• Yang J, Liu H, Sun H et al. Construction of induced pluripotent stem cell line (ZZUi0017-A) from the fibroblast cells of a female patient with CACNA1A mutation by unintegrated reprogramming approach Stem Cell Res 2020-08-04 [PMID: 32791484] (ICC/IF, Human)
• Li W, Zhang N, Jin C et al. MUC1-C drives stemness in progression of colitis to colorectal cancer JCI Insight 2020-05-19 [PMID: 32427590] (KO, WB, Mouse)
• Lu H, Xie Y, Tran L et al. Chemotherapy-induced S100A10 recruits KDM6A to facilitate OCT4-mediated breast cancer stemness J Clin Invest. 2020-05-19 [PMID: 32427586] (WB, KD, Human)
• Yang T, Qin J, Zhang Q et Al. Generation of induced pluripotent stem cell line (ZZUi0018-A ) from a patient with spinocerebellar ataxia type 6 Stem Cell Res. [PMID: 32229428] (FLOW, ICC/IF, Human)
• Gao Y, Wang Y, Wang Z et al. Generation of induced pluripotent stem cell line (ZZUi0016-A) from dermal fibroblasts of a normal human Stem Cell Res. [PMID: 32007761] (FLOW, ICC/IF, Human)
• Sun J, Zhang X, Cong Q, Wang C et Al. Generation of refractory schizophrenia patient-derived induced pluripotent stem cell line UJSi001-A Stem Cell Res 2019-05-19 [PMID: 31102833]
• Sohni A, Tan K, Song HW et al. The Neonatal and Adult Human Testis Defined at the Single-Cell Level Cell Rep 2019-02-05 [PMID: 30726734] (IF/IHC, Human)
• Han YM, Bedarida T, Ding Y et al. B-Hydroxybutyrate Prevents Vascular Senescence through hnRNP A1-Mediated Upregulation of Oct4. Mol. Cell 2018-09-20 [PMID: 30197300] (ICC/IF, Mouse)
• Cheng CC, Chang J, Huang SC et al. YM155 as an inhibitor of cancer stemness simultaneously inhibits autophosphorylation of epidermal growth factor receptor and G9a-mediated stemness in lung cancer cells PLoS ONE 2017-08-08 [PMID: 28787001] (WB, Human)
• McCulloh Christopher J, Olson Jacob K, Zhou Yu et al. Stem cells and necrotizing enterocolitis: A direct comparison of the efficacy of multiple types of stem cells. Journal of Pediatric Surgery 2018-06-01 [PMID: 28366560] (FLOW, Rat)
• Lupicka M, Socha B, Szczepanska A, Korzekwa A. Expression of pluripotency markers in the bovine uterus with adenomyosis. Reprod Biol Endocrinol 2015-01-01 [PMID: 26416515] (WB)
• Lin HY, Lee DC, Wang HD et al. Activation of FGF1B Promoter and FGF1 Are Involved in Cardiogenesis Through the Signaling of PKC, but Not MAPK. Stem Cells Dev 2015-12-15 [PMID: 26414172]
• Lu H Samanta D Xiang L et al. Chemotherapy triggers HIF-1-dependent glutathione synthesis and copper chelation that induces the breast cancer stem cell phenotype. Proc Natl Acad Sci U S A. 2015-07-30 [PMID: 26229077] (WB, Human)
• Darekar SD, Mushtaq M, Gurrapu S et al. Mitochondrial ribosomal protein S18-2 evokes chromosomal instability and transforms primary rat skin fibroblasts. Oncotarget. 2015-05-12 [PMID: 26023799] (WB, Rat)
  
  Details:
  OCT4 antibody was used for WB analysis of lysates from primary rat skin fibroblasts /RSFs and immortalized cells namely clone 17, clone 13 and clone 6. Oct4 was found to be induced in all clones of immortalized RSFs compared with the parental cells (Figure 2d).
• Liu JW, Hsu YC, Kao CY et al. LIF-induced Stat3 signaling suppresses FGF1-induced Erk1/2 activation to inhibit the downstream differentiation in mouse embryonic stem cells Stem Cells Dev 2012-12-04 [PMID: 23205673]
• Ruggeri RR, Watanabe Y, Meirelles F et al. The use of parthenotegenetic and IVF bovine blastocysts as a model for the creation of human embryonic stem cells under defined conditions J Assist Reprod Genet 2012-09-29 [PMID: 23054358] (ICC/IF, Bovine)
• Cho YM, Kim YS, Kang MJ et al. Long-term recovery of irradiated prostate cancer increases cancer stem cells Prostate 2012-12-01 [PMID: 22513891] (WB, Human)
• Yenamandra SP, Darekar SD, Kashuba V et al. Stem cell gene expression in MRPS18-2-immortalized rat embryonic fibroblasts. Cell Death Dis 2012-01-01 [PMID: 22258410] (WB, Rat)

Reviews for OCT4 Antibody (NB100-2379) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Human.

reviewed by: Verified Customer WB Human 05/18/2020

Summary

• Application: Western Blot
• Sample Tested: Breast cancer cells
• Species: Human
• Lot: C-3

reviewed by: Verified Customer WB Human 10/11/2018

Summary

• Application: Western Blot
• Sample Tested: human head and neck cancer cell lines
• Species: Human
• Lot: C-3

FAQs for OCT4 Antibody (NB100-2379). (Showing 1 - 1 of 1 FAQs).

1. We are looking for the following markers to use to stain cells derived from both human and mouse embryonic and induced pluripotent stem cells: undifferentiated cells, neurons, oligodendrocytes, and astrocytes.
   • For ES markers some popular targets are OCT4 (NB100-2379), KLF4 (NBP2-17070) and SOX2 (NB110-37235). PEA15 is expressed in Astrocytes and this might be one of interest for you (NBP1-61390). OCT4 is one that can also be expressed in Oligodendrocytes.

Bioinformatics

• Gene Symbol: POU5F1
• Entrez:
  • 5460(Human)
  • 18999(Mouse)
• Uniprot:
  • Q01860()