Overview
- Product nameAnti-RNF20 antibodySee all RNF20 primary antibodies ...
- DescriptionRabbit polyclonal to RNF20
- Tested applicationsWB, ICC/IF, IHC-P, IP more details
- Species reactivityReacts with: Chicken, Human
Predicted to work with: Mouse - Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human RNF20.
(Peptide available as ab32628.)
- Positive controlIn Western blot, this antibody gave a positive signal in chicken liver tissue lysate as well as in the following whole cell lysates: HeLa; Jurkat; A431; HEK293; MCF7; U20S. This antibody also gave a positive signal in HeLa whole cell lysates in Immunofluorescence and Human Testis tissue lysate in Immunohistochemistry.
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab32629 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 114 kDa).Can be blocked with RNF20 peptide (ab32628). |
| ICC/IF | ICC/IF: Use a concentration of 1 µg/ml. |
| IHC-P | IHC-P: 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| IP | IP: Use at an assay dependent concentration. |
Target
- RelevanceRNF20 is an E3 ubiquitin ligase protein that mediates monoubiquitination of histone H2B and the methylation of histone H3. It forms a ubiquitin ligase complex in cooperation with the E2 enzyme UBE2E1/UBCH6. It thereby plays a central role in histone code and gene regulation. It is required for transcriptional activation of Hox genes. It is recruited to the MDM2 promoter, probably by being recruited by p53/TP53, and thereby acts as a transcriptional coactivator.
- Cellular localizationNuclear
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Database links
- Entrez Gene: 427310 Chicken
- Entrez Gene: 56254 Human
- Entrez Gene: 56254 Human
- Entrez Gene: 109331 Mouse
- Omim: 607699 Human
- SwissProt: Q5ZLS3 Chicken
- SwissProt: Q5VTR2 Human
- SwissProt: Q5VTR2 Human
- SwissProt: Q7TT11 Mouse
- Unigene: 656088 Human
- Unigene: 24765 Mouse
see all
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Alternative names
- BRE1 A antibodyBRE1 antibodyBRE1 E3 ubiquitin ligase homolog antibody
- BRE1A antibodyE3 ubiquitin-protein ligase BRE1A antibodyhBRE1 antibodyHomolog of S. cerevisiae BRE1 antibodyRING finger protein 20 antibodyRing finger protein 20 E3 ubiquitin protein ligase antibody
see all
Anti-RNF20 antibody images
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Western blot - RNF20 antibody (ab32629)All lanes : Anti-RNF20 antibody (ab32629) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 :A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
Lane 4 :HEK293 Human embryonic kidney cell line Whole Cell Lysate (ab7902)
Lane 5 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
Performed under reducing conditions.
Predicted band size : 114 kDa
Observed band size : 120 kDa (why is the actual band size different from the predicted?) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - RNF20 antibody (ab32629)Image courtesy of Human Protein Atlas
ab32629 staining RNF20 in human testis, showing a distinct and strong staining pattern in ductus seminiferus cells. Paraffin embedded human skin tissue was incubated with ab32629 (1/50 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
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ab32629 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org -
Immunocytochemistry/ Immunofluorescence - RNF20 antibody (ab32629)This image is courtesy of an Abreview submitted by Dr Kirk McManusab32629 (1/500) staining RNF20 in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI to highlight the nucleus (red). Please refer to abreview for further experimental details. -
Immunocytochemistry/ Immunofluorescence - RNF20 antibody (ab32629)ICC/IF image of ab32629 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32629, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml. -
Western blot - Anti-RNF20 antibody (ab32629)All lanes : Anti-RNF20 antibody (ab32629) at 1/1000 dilution
Lane 1 : Whole cell lysate prepared from Ntera2 cells
Lane 2 : Whole cell lysate prepared from Ntera2 cells + siRNA against RNF20 for 48 hours
Lysates/proteins at 20 µg per lane.
Secondary
HRP conjugated goat polyclonal at 1/10000 dilution
developed using the ECL technique
Predicted band size : 114 kDa
Observed band size : 120 kDa (why is the actual band size different from the predicted?)
Exposure time : 20 secondsImage courtesy of an anonymous Abreview.
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Western blot - Anti-RNF20 antibody (ab32629)Anti-RNF20 antibody (ab32629) at 1 µg/ml + Liver (Chicken) Tissue Lysate at 10 µg
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 114 kDa
Observed band size : 140 kDa (why is the actual band size different from the predicted?)
Exposure time : 20 minutes
References for Anti-RNF20 antibody (ab32629)
This product has been referenced in:
- Vethantham V et al. Dynamic loss of H2B ubiquitylation without corresponding changes in H3K4 trimethylation during myogenic differentiation. Mol Cell Biol 32:1044-55 (2012). ChIP . Read more (PubMed: 22252316) »
- Gao Z et al. Resveratrol induces cellular senescence with attenuated mono-ubiquitination of histone H2B in glioma cells. Biochem Biophys Res Commun 407:271-6 (2011). Human . Read more (PubMed: 21481687) »
