Ubiquitin Antibody (Ubi-1) - BSA Free

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Ubiquitination of PML in response to As2O3. PML-/- MEFs were transiently transfected with the empty vector (EV) or PMLIII (left panel) or contructs encoding PMLIII, PMLIII-SIM or PMLVI (right panel). Two days later, they were treated with As2O3 for 1 h. The cell extracts were immunoprecipitated with rabbit anti-PML antibody and the immunoprecipitate was analyzed by Western blot using anti-ubiquitin and anti-PML antibodies. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0044949) licensed under a CC-BY license.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Ubiquitination of PML in response to As2O3. PML-/- MEFs were transiently transfected with the empty vector (EV) or PMLIII (left panel) or contructs encoding PMLIII, PMLIII-SIM or PMLVI (right panel). Two days later, they were treated with As2O3 for 1 h. The cell extracts were immunoprecipitated with rabbit anti-PML antibody and the immunoprecipitate was analyzed by Western blot using anti-ubiquitin and anti-PML antibodies. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0044949) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Ubiquitin Antibody (Ubi-1) [NB300-130] - Study of mutant BRAF protein found within the inclusions. BRAFV600E/ubiquitin double-IF labeling. A nucleus with a large inclusion is shown with positive immunoreactivity for both mutant BRAF (red staining) and ubiquitin (green staining); the merged color yellow (arrow) points out the co-localisation of BRAFV600E and ubiquitin in the same inclusion. Image collected and cropped by CiteAb from the following publication (////doi.org/10.1371/journal.pone.0226199) licensed under a CC-BY license.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Analysis of HEK293 cell lysates using mouse mAb Ubiquitin, dilution 1:1000 (Green). [1] protein standard (Red), [2] cells maintained in normal medium, [3] cells treated with 10uM of proteasome inhibitor lactacystin (Lc) for 16hrs. Lysed cells were electrophoresed on 4-20% SDS-PAGE and transferred to PVDF membranes. The smear detected above the 200kDa standard represent accumulation of ubiquitinated proteins in proteasome inhibitor-Lc treated cells. The prominent band at 8kDa corresponds to monoubiquitin. Rabbit pAb to HSP60, dilution 1:5000 (Red) was used as a loading control.

Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - FFPE section of cerebral cortex of an Alzheimer patient processed with Ubiquitin antibody using HRP/DAB (Brown) and also stained with haemotoxylin (Blue). A typical flame shaped tangle is seen in a pyramidal neuron in the center and is surrounded by some dystrophic neurites, also strongly ubiquitin positive. Both are commonly seen in cortical and hippocampal Alzheimer brain sections and are typical for this disease, but are rare or absent in healthy brain.

Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - Staining of Ubiquitin in hippocampal tissue from an Alzheimer patient.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Molecular Analysis of XopD Ub/Ubl Specificity. S. lycopersicum protein extract blotted for total Ub (Ubiquitin; Novus Biologicals) following a 1 hr room temperature treatment with the XopD variants used in (K) at 1 uM final concentration. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27425412/) licensed under a CC-BY license.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Western blot of Ubiquitin expression on (Lane 1) poly-ubiquitin (lys63 linked), (Lane 2) pure ubiquitin and crude homogenates of adult rat, (Lane 3) cortex, (Lane 4) cerebellum and (Lane 5) brain stem.

Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - IHC staining of Ubiquitin in human rectal cancer using DAB with hematoxylin counterstain.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - In Vivo Applications of Affimers(A) HA-NleL 293 T-Rex cells were induced with 1 μg/mL doxycycline for 12 hr or left untreated. Whole-cell lysate (WCL) blots are shown for actin, HA(-NleL), Ub, & K6 chains. Western blots with the K6 affimer are also shown after Ub enrichment using TUBEs.(B) TUBE-PD of HEK293 cells after 1 hr of MG132 (10 μM) without further treatment (−) or with additional UV (40 J/m2) or CCCP treatment (10 μM for 1 hr) & subsequently blotted with the K6 affimer. Input controls are shown for total Ub & actin & gamma H2AX. The relative signal increase from two experiments is shown below the respective lanes.(C) Expression of WT or catalytically inactive (C431S) Parkin in HeLa Flp-In cells was induced with 0.2 μg/mL doxycycline for 16 hr. Mitochondria were depolarized with O/A for 2 hr. WCL inputs are shown for total Ub, expressed Parkin, TOM20, & actin. The TUBE-PD was also blotted using the K6 affimer & total Ub.(D) Confocal fluorescence microscopy images of cells as in (B) stained with K6 affimer (green), TOM20 (red), & DAPI (blue). Cells & a magnified area are outlined in white. Scale bars correspond to 20 μm.See also Figure S5. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28943312), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - In Vivo Applications of Affimers(A) HA-NleL 293 T-Rex cells were induced with 1 μg/mL doxycycline for 12 hr or left untreated. Whole-cell lysate (WCL) blots are shown for actin, HA(-NleL), Ub, & K6 chains. Western blots with the K6 affimer are also shown after Ub enrichment using TUBEs.(B) TUBE-PD of HEK293 cells after 1 hr of MG132 (10 μM) without further treatment (−) or with additional UV (40 J/m2) or CCCP treatment (10 μM for 1 hr) & subsequently blotted with the K6 affimer. Input controls are shown for total Ub & actin & gamma H2AX. The relative signal increase from two experiments is shown below the respective lanes.(C) Expression of WT or catalytically inactive (C431S) Parkin in HeLa Flp-In cells was induced with 0.2 μg/mL doxycycline for 16 hr. Mitochondria were depolarized with O/A for 2 hr. WCL inputs are shown for total Ub, expressed Parkin, TOM20, & actin. The TUBE-PD was also blotted using the K6 affimer & total Ub.(D) Confocal fluorescence microscopy images of cells as in (B) stained with K6 affimer (green), TOM20 (red), & DAPI (blue). Cells & a magnified area are outlined in white. Scale bars correspond to 20 μm.See also Figure S5. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28943312), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - HUWE1 Assembles K6 Chains In Vitro & In Vivo(A) Table summarizing proteins identified with the corresponding number peptide-spectrum matches (PSMs) in three replicates of DUB-treated K6 affimer pull-downs.(B) In vitro assembly reaction of the HECT E3 HUWE1 with Ub WT, Ub K6R, Ub K11R, & Ub K48R on Coomassie. Arrows indicate K6 diUb.(C) Linkage composition of HUWE1-generated diUb after 1 hr as determined by AQUA MS.(D) AQUA-derived total cellular chain composition of HUWE1−/− HeLa & parental cells after TUBE-based enrichment. Error bars indicate mean ± SD from n = 3. ∗p < 0.05, according to a two-tailed Student’s t test. N.S., not significant.(E) TUBE-PD from a doxycycline-inducible HUWE1 shRNA Ls174T cell line blotted with the K6-specific affimer, with input controls for actin, total Ub, & HUWE1.(F) K6 affimer pull-down in doxycycline-inducible HUWE1 shRNA Ls174T cells blotted against Mfn2. Cells were left untreated or treated with 10 μg/mL MG132 for 4 hr and/or 1 μg/mL doxycycline for 72 hr. Pull-downs were incubated with 250 nM USP21 as indicated, with a K48 blot to show completeness of the deubiquitination reaction. Input controls are shown for Mfn2, actin, & HUWE1.See also Figure S7. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28943312), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - In Vivo Applications of Affimers(A) HA-NleL 293 T-Rex cells were induced with 1 μg/mL doxycycline for 12 hr or left untreated. Whole-cell lysate (WCL) blots are shown for actin, HA(-NleL), Ub, & K6 chains. Western blots with the K6 affimer are also shown after Ub enrichment using TUBEs.(B) TUBE-PD of HEK293 cells after 1 hr of MG132 (10 μM) without further treatment (−) or with additional UV (40 J/m2) or CCCP treatment (10 μM for 1 hr) & subsequently blotted with the K6 affimer. Input controls are shown for total Ub & actin & gamma H2AX. The relative signal increase from two experiments is shown below the respective lanes.(C) Expression of WT or catalytically inactive (C431S) Parkin in HeLa Flp-In cells was induced with 0.2 μg/mL doxycycline for 16 hr. Mitochondria were depolarized with O/A for 2 hr. WCL inputs are shown for total Ub, expressed Parkin, TOM20, & actin. The TUBE-PD was also blotted using the K6 affimer & total Ub.(D) Confocal fluorescence microscopy images of cells as in (B) stained with K6 affimer (green), TOM20 (red), & DAPI (blue). Cells & a magnified area are outlined in white. Scale bars correspond to 20 μm.See also Figure S5. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28943312), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt, Po, Bv, Ce, Ch, Dr, Eq, Pl, Pm, Ze
• Applications: WB, ICC/IF, IHC, WB
• Clone: Ubi-1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

Ubiquitin Antibody (Ubi-1) - BSA Free Summary

• Immunogen: Purified bovine erythrocyte ubiquitin cross-linked with glutaraldehyde to KLH
• Specificity: Recognizes polyubiquitin chains much more strongly than monoubiquitinated molecules or free ubiquitin. Specifically recognizes ubiquitinated cytoplasmic inclusion bodies.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: RPS27A
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • Immunoblotting
  • Immunocytochemistry/ Immunofluorescence 1:2000
  • Immunohistochemistry 1:2000
  • Immunohistochemistry-Frozen 1:2000
  • Immunohistochemistry-Paraffin 1:2000
  • Western Blot 1:1000-1:2000
• Application Notes: This Ubiquitin Antibody (Ubi-1) is useful for Western blot, ELISA, Immunocytochemistry/Immunofluorescence, and Immunohistochemistry on both paraffin-embedded and frozen sections. In Western blot a band can be seen at 8.5 kDa. This antibody is excellent for the detection of ubiquitinated inclusions seen in human neurodegenerative diseases such as the neurofibrillary tangles of Alzheimer's disease. Use in immunoblotting reported in scientific literature (PMID: 31727601).
• Theoretical MW: 8.5 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.

Reactivity Notes

Plant reactivity reported in scientific literature (PMID: 19147500).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 50% PBS, 50% glycerol
• Preservative: 0.035% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Affinity purified

Alternate Names for Ubiquitin Antibody (Ubi-1) - BSA Free

• RPS27A
• UBA52
• UBB ubiquitin B
• UBB
• UBC
• Ubiquitin

Background

Ubiquitin is a highly conserved protein of about 8.5 kDa molecular weight, which has a normal role in the targeting of proteins for proteolytic degradation. To perform this function, the protein to be degraded is first covalently attached to the C-terminus of ubiquitin, and the ubiquitinated complex is then recognized by a complex of degradative enzymes. Interestingly, ubiquitin also becomes covalently bonded to many types of pathological inclusions, which appear to be resistant to normal degradation. Therefore, ubiquitin antibodies are very useful for studies of these inclusions. For example, the neurofibrillary tangles and paired helical filaments diagnostic of Alzheimer's disease, Lewy bodies seen in Parkinson's disease, and Pick bodies found in Pick's disease are all heavily ubiquitinated and can be readily visualized with ubiquitin antibodies.

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 Ubiquitin Antibody (NB300-130)(52)

Publications using NB300-130

• Hamamoto K, Liang X, Ito A et Al. Unveiling the physiological impact of ESCRT-dependent autophagosome closure by targeting the VPS37A ubiquitin E2 variant-like domain Cell Rep 2024-11-27 [PMID: 39607828]
• Volik PI, Zamaraev AV, Egorshina AY et Al. Ally or traitor: the dual role of p62 in caspase-2 regulation Cell Death Dis 2024-11-14 [PMID: 39543123]
• Muhammad T, Edwards SL, Morphis AC et al. Non-cell-autonomous regulation of germline proteostasis by insulin/IGF-1 signaling-induced dietary peptide uptake via PEPT-1 The EMBO Journal 2024-09-16 [PMID: 39284915]
• Michel MA, Scutts S, Komander D., et Al. Secondary interactions in ubiquitin-binding domains achieve linkage or substrate specificity Cell Rep 2024-07-25 [PMID: 39052481]
• Qingpeng Xie, Bin Hu, Haosong Li Acetylation- and ubiquitination-regulated SFMBT2 acts as a tumor suppressor in clear cell renal cell carcinoma Biology Direct 2024-05-11 [PMID: 38734627]
• Hackerova L, Klusackova B, Zigo M et al. Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation Frontiers in veterinary science 2023-03-23 [PMID: 37035827] (FLOW, Porcine)
  
  Details:
  Duroc boars sperm; dilution: 1:10
• Yang S, Xiang J, Ma C et al. Sp1-like protein KLF13 acts as a negative feedback regulator of TGF-? signaling and fibrosis Cell reports 2023-04-07 [PMID: 37029927]
• Yonenobu Y, Beck G, Kido K et al. Neuropathology of spinocerebellar ataxia type 8: Common features and unique tauopathy Neuropathology : official journal of the Japanese Society of Neuropathology 2023-01-26 [PMID: 36703300]
• Guo Y, Chomiak AA, Hong Y et al. Histone H2A ubiquitination resulting from Brap loss of function connects multiple aging hallmarks and accelerates neurodegeneration iScience 2022-07-15 [PMID: 35754718]
• Chomiak AA, Guo Y, Kopsidas CA et al. Nde1 is required for heterochromatin compaction and stability in neocortical neurons iScience 2022-06-17 [PMID: 35601919] (Western Blot, Human)
• Kwon M, Oh T, Jang M et al. Kurarinone induced p53-Independent G0/G1 cell cycle arrest by degradation of K-RAS via WDR76 in human colorectal cancer cells European Journal of Pharmacology 2022-04-01 [PMID: 35381263] (WB, Human)
• Mohan AK Regulation of Inflammatory Signalling by Caspases and M1-linked Ubiquitin Chains in Drosophila melanogaster Thesis 2023-01-01 [PMID: 35263507]
• Zhang C, Tang Q, Xia H et al. PKM2 compensates for proteasome dysfunction by mediating the formation of the CHIP-HSP70-BAG3 complex and the aggregation of ubiquitinated proteins FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2022-01-01 [PMID: 34951719] (ICC/IF, Human)
• Ito Y, Yamamoto T, Miyai K et al. Ascorbic acid-2 glucoside mitigates intestinal damage during pelvic radiotherapy in a rat bladder tumor model International journal of radiation biology 2021-12-06 [PMID: 34871138]
• 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]
• Jo J, Yang L, Tran HD et al. Lewy-body Like Inclusions in Human Midbrain Organoid Carrying Glucocerebrosidase and Alpha Synuclein Mutations Annals of neurology 2021-07-20 [PMID: 34288055] (IHC-P)
• Robichaud S, Fairman G, Vijithakumar V et al. Identification of novel lipid droplet factors that regulate lipophagy and cholesterol efflux in macrophage foam cells Autophagy 2021-02-26 [PMID: 33590792] (Western Blot, Human)
• Schwertheim S, KAlsch J, Jastrow H et al. Characterization of two types of intranuclear hepatocellular inclusions in NAFLD Sci Rep 2020-10-06 [PMID: 33024131] (ICC/IF, Human)
• Thurner L, Hartmann S, Bewarder M et al. Identification of the atypically modified autoantigen Ars2 as the target of B-cell receptors from activated B cell-type diffuse large B-cell lymphoma Haematologica 2020-07-16 [PMID: 32675228] (Human)
• Chen Z, Wasney G A et al. Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12. Open Biol 2020-01-06 [PMID: 32574548] (WB, Human)
• Yoon YM, Lee JH, Song KH et al. Melatonin-stimulated exosomes enhance the regenerative potential of chronic kidney disease-derived mesenchymal stem/stromal cells via cellular prion proteins J. Pineal Res. 2020-01-27 [PMID: 31989677]
• Schultz E, Jones T, Clamp H, Barr K New insights into intranuclear inclusions in thyroid carcinoma: Association with autophagy and with BRAFV600E mutation PLoS ONE 2019-12-16 [PMID: 31841566] (ICC/IF, Human)
• Sn S, Pandurangi J, Murumalla R et al. Small molecule modulator of aggrephagy regulates neuroinflammation to curb pathogenesis of neurodegeneration EBioMedicine 2019-11-11 [PMID: 31727601] (IB, Human)
• Swatek KN, Usher JL, Kueck AF et al. Insights into ubiquitin chain architecture using Ub-clipping Nature 2019-08-01 [PMID: 31413367] (WB, Human)
• RB Damgaard, PR Elliott, KN Swatek, ER Maher, P Stepensky, O Elpeleg, D Komander, Y Berkun OTULIN deficiency in ORAS causes cell type-specific LUBAC degradation, dysregulated TNF signalling and cell death EMBO Mol Med, 2019-03-01;11(3):. 2019-03-01 [PMID: 30804083]
• Li L, Guturi KKN, Gautreau B et al. Ubiquitin ligase RNF8 suppresses Notch signaling to regulate mammary development and tumorigenesis. J. Clin. Invest. 2018-10-01 [PMID: 30222135]
• Aalto AL, Mohan AK, Schwintzer L et al. M1-linked ubiquitination by LUBEL is required for inflammatory responses to oral infection in Drosophila. Cell Death Differ. 2018-07-19 [PMID: 30026495] (WB, Drosophila)
• Gladkova C, Maslen SL, Skehel JM, Komander D Mechanism of Parkin activation by PINK1 Nature. 2018-06-05 [PMID: 29995846] (WB)
• Gogoi M, Ravikumar V, Dixit NM, Chakravortty D. Salmonella escapes K63 ubiquitination mediated MHC II endosomal proteolysis via modulation of endosomal acidification in dendritic cells. Pathog Dis 2017-12-26 [PMID: 29293966] (WB, ICC/IF)
• Deger SM, Hung AM, Gamboa JL et al. Systemic inflammation is associated with exaggerated skeletal muscle protein catabolism in maintenance hemodialysis patients JCI Insight 2017-11-16 [PMID: 29202452] (WB, Mouse)
• Gersch M, Gladkova C, Schubert AF et al. Mechanism and regulation of the Lys6-selective deubiquitinase USP30. Nat. Struct. Mol. Biol. 2017-09-25 [PMID: 28945249] (WB, Human)
• Michel MA, Swatek KN, Hospenthal MK, Komander D. Ubiquitin Linkage-Specific Affimers Reveal Insights into K6-Linked Ubiquitin Signaling Mol. Cell 2017-09-20 [PMID: 28943312] (WB, Human)
• Kaul Z, Chakrabarti O. Tumor susceptibility gene 101, TSG101, REGULATES predisposition towards apoptosis VIA ESCRT machinery accessory Proteins. Mol. Biol. Cell 2017-05-24 [PMID: 28539405] (WB, IF/IHC, Human)
• Damgaard RB, Walker JA, Marco-Casanova P et al. The Deubiquitinase OTULIN Is an Essential Negative Regulator of Inflammation and Autoimmunity. Cell. 2016-08-25 [PMID: 27523608] (WB)
• Begum G, Yan HQ, Li L et al. Docosahexaenoic Acid Reduces ER Stress and Abnormal Protein Accumulation and Improves Neuronal Function Following Traumatic Brain Injury. J. Neurosci. 2014-03-05 [PMID: 24599472] (IHC-Fr, Rat)
• Ejlerskov P, Rasmussen I, Nielsen TT et al. Tubulin Polymerization Promoting Protein (TPPP/p25alpha) promotes unconventional secretion of alpha-synuclein through exophagy by impairing autophagosome-lysosome fusion. J Biol Chem 2013-04-29 [PMID: 23629650] (ICC/IF, Rat)
• Rosenfeldt MT, Nixon C, Liu E et al. Analysis of macroautophagy by immunohistochemistry Autophagy 2012-06-01 [PMID: 22562096] (IF/IHC, Mouse)
• Guo J Y, Chen H Y et al. Activated Ras requires autophagy to maintain oxidative metabolism and tumorigenesis. Genes Dev 2011-01-03 [PMID: 21317241] (IF/IHC, Mouse)
• Stuttmann J, Lechner E, Guerois R et al. COP9 Signalosome- 26S Proteasome-dependent Regulation of SCFTIR1 Accumulation in Arabidopsis. J Biol Chem;284(12):7920-7930. 2009-01-01 [PMID: 19147500] (WB, Plant)
• Dayal S, Sparks A, Jacob J et al. Suppression of the Deubiquitinating Enzyme USP5 Causes the Accumulation of Unanchored Polyubiquitin the Activation of p53. J Biol Chem;284(8):5030-41. 2009-02-20 [PMID: 19098288] (WB, Human)
• Watson MR, Lagow RD, Xu K et al. A Drosophila Model for Amyotrophic Lateral Sclerosis Reveals Motor Neuron Damage by Human SOD1. J Biol Chem;283(36):24972-24981. 2008-01-01 [PMID: 18596033] (IF/IHC, Drosophila)
• Wallace I, Baek K, Prabu J et al. Insights into the ISG15 transfer cascade by the UBE1L activating enzyme bioRxiv 2023-04-06
• Pruneda JN, Durkin CH, Geurink PP et al. The Molecular Basis for Ubiquitin and Ubiquitin-like Specificities in Bacterial Effector Proteases Mol. Cell 2016-07-21 [PMID: 27425412] (WB)
• Schwab AJ, Ebert AD. Neurite Aggregation and Calcium Dysfunction in iPSC-Derived Sensory Neurons with Parkinson's Disease-Related LRRK2 G2019S Mutation. Stem Cell Reports. 2015-12-08 [PMID: 26651604] (ICC/IF, Human)
• Maroui MA, Kheddache-Atmane S, El Asmi F et al. Requirement of PML SUMO Interacting Motif for RNF4- or Arsenic Trioxide-Induced Degradation of Nuclear PML Isoforms PLoS One 2012-01-01 [PMID: 23028697] (WB, Human)
• Sun K, Johnson B S et al. Mitochondrial dysfunction precedes neurodegeneration in mahogunin (Mgrn1) mutant mice. Neurobiol Aging 2007-01-12 [PMID: 17720281] (WB, Mouse)
• Ryazantsev S, Yu W H et al. Lysosomal accumulation of SCMAS (subunit c of mitochondrial ATP synthase) in neurons of the mouse model of mucopolysaccharidosis III B. Mol Genet Metab 2007-01-04 [PMID: 17185018] (ICC/IF, Mouse)
• Hawryluk M J, Keyel P A et al. Epsin 1 is a polyubiquitin-selective clathrin-associated sorting protein. Traffic 2006-01-03 [PMID: 16497222] (WB, Human)
• Chang L, Kamata H, Solinas G et al. The E3 ubiquitin ligase itch couples JNK activation to TNFalpha-induced cell death by inducing c-FLIP(L) turnover. Cell. 2006-02-10 [PMID: 16469705] (WB)
  
  Details:
  WB: 1. IkBa [IMG-127A], (wt hepatocytes incubated with proteasome inhibitor MG-132 and TNFa or CHX added), Fig. 3a 2. Ubiquitin [IMG-5021]: (wt and Jnk1-/- injected mouse liver with ConA), Fig. 3b (wt mouse liver injected with LPS and Cal-N and JNk-inhibi
• Cripps, D et al. Alzheimer disease-specific conformation of hyperphosphorylated paired helical filament-tau is polyubiquitinated through Lys-438, Lys-11, Lys-6 ubiquitin conjugation. J Biol Chem; 281: 10825-10838. 2006-01-01 [PMID: 16443603] (WB, Human)
• Lichtenfels, R et al. Identification of metabolic enzymes in renal cell carcinoma utilizing PROTEOMEX analyses. Biochimica et Biophysica Acta; 36772:1-11. 2002-01-01 [PMID: 12637008] (WB, Human)
• Zhang H, Wang T, Liu H et al. Boost therapy of hepatocellular carcinoma by amplifying vicious cycle between mitochondrial oxidative stress and endoplasmic reticulum stress via biodegradable ultrasmall nanoparticles and old drug Nano Today 2022-10-01 (WB, ICC/IF, Human)
  
  Details:
  ICC/If Dilution: 1:200; WB Dilution: 1:1000

Reviews for Ubiquitin Antibody (NB300-130) (4)

Average Rating: 4.3

(Based on 4 reviews)

We have 4 reviews tested in 3 species: Human, Mouse, Human and Mouse.

reviewed by: Verified Customer IHC-P Human 08/25/2021

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Adult heart
• Species: Human
• Lot: Ubi-1

reviewed by: Verified Customer WB Mouse 03/27/2018

Summary

• Application: Western Blot
• Sample Tested: Mouse skeletal muscle homogenate
• Species: Mouse
• Lot: 1027092-3

Comments

• Comments: Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130SS] - Total protein from Mouse skeletal muscle homogenate, separated on a 4-12% gel by SDS-PAGE, transferred to nitrocellulose membrane and blocked in 5% non-fat milk for 1h at room temperature. The membrane was probed with anti-Myogenin 1:1000 in non-fat milk.

reviewed by: Rune Damgaard WB Human and Mouse 08/04/2017

Summary

• Application: Western Blot
• Sample Tested: 293 whole cell lysate,HCT116 whole cell lysate,Hela whole cell lysate,MEF whole cell lysate,mouse bone marrow-derived macrophage cells,mouse liver lysate
• Species: Human and Mouse
• Lot: 1027092-2

Comments

• Comments: Works well for WB on total cell lysates (boiling the membrane (nitrocellulose) can increase signal drastically if it is low, e.g. when blotting on primary cells/tissues), and it works really well for WB on Ubiquitin pulldowns, e.g. after a TUBE (tandem ubiquitin-binding entity) pulldown.

reviewed by: Verified Customer IF Mouse 05/27/2009

Summary

• Application: Immunofluorescence
• Sample Tested: primary mouse neuronal culture
• Species: Mouse
• Lot: 120103

FAQs for Ubiquitin Antibody (NB300-130). (Showing 1 - 1 of 1 FAQs).

1. Regarding the WB image on datasheet of NCX1 antibody (6H2) (NB300-127) , can you confirm if lane 2 is a rabbit heart sample? In addition, can you provide the dilution fold and loading mount used to obtain this image? Lastly, would you happen to know if t  ..show answer..
   • A protein stabilizer has not been added to the antibody NB300-130. However, note that this antibody is supplied as ascites fluid, not as a purified antibody. If you are not familiar with this format, the following descriptions will be useful: Ascites fluid is a complex solution containing multiple proteins and background antibodies as well as the antibody of interest. Ascites contains the antibody produced by the hybridoma cells as well as many other proteins, including background antibodies produced by the mice and albumin. Antibody concentrations in ascites are typically high, but an accurate concentration of the target antibody cannot be readily determined. Therefore, unfortunately you cannot determine the concentration of the ubiquitin antibody within NB300-130 as sold. This is why this product's pack sizes are volumes rather than masses, and why we state on our datasheet: 'This product is unpurified. The exact concentration of antibody is not quantifiable.' All I can suggest is that you carry out antigen affinity chromatography of the antibody, and then determine its concentration.

Bioinformatics

• Gene Symbol: RPS27A
• Entrez:
  • 7314(Human)
  • 7316(Human)
• Uniprot:
  • P0CG47()
  • P0CG48(Human)