Overview

  • Product nameAnti-RNF20 antibodySee all RNF20 primary antibodies ...
  • Description
    Rabbit polyclonal to RNF20
  • Tested applicationsWB, ICC/IF, IHC-P, IP more details
  • Species reactivity
    Reacts 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 control
    • In 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

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 Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 114 kDa).Can be blocked with Human RNF20 peptide (ab32628).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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
  • Database links
  • Alternative names
    • BRE1 A antibody
    • BRE1 antibody
    • BRE1 E3 ubiquitin ligase homolog antibody
    • BRE1A antibody
    • E3 ubiquitin-protein ligase BRE1A antibody
    • hBRE1 antibody
    • Homolog of S. cerevisiae BRE1 antibody
    • RING finger protein 20 antibody
    • Ring finger protein 20 E3 ubiquitin protein ligase antibody
    see all

Anti-RNF20 antibody images

  • 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?)
  • 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.

    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

    .
  • ab32629 (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.

    See Abreview

  • 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.
  • 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 seconds

    Image courtesy of an anonymous Abreview.

    See Abreview

  • Anti-RNF20 antibody (ab32629) at 1 µg/ml + Liver (Chicken) Tissue Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (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) »

See all 4 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Sample Human Cell (Human primary fibroblasts)
Specification Human primary fibroblasts
Permeabilization Yes - 0.2% Triton-X100
Fixative Formaldehyde
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Abcam user community

Verified customer

Submitted Dec 12 2013

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (12%)
Sample Human Cell lysate - whole cell (U2OS)
Specification U2OS
Blocking step Milk as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 23°C
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Dr. Abigail Buchwalter

Verified customer

Submitted Sep 30 2013

I am sorry to hear that your customer has been experiencing problems using this product in Western blot application.

The Western blot form attached to the e-mail has not been completed at all, it is empty. Though, I have successfully received...

Read More
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse lung)
Loading amount 100000 cells
Specification Mouse lung
Gel Running Conditions Reduced Denaturing (23.5%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Dec 03 2012

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (Breast cell line)
Total protein in input 100000 cells
Specification Breast cell line
Immuno-precipitation step Protein A/G
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Submitted Nov 28 2012

Application Western blot
Sample Human Cell lysate - whole cell (MCF10A- Breast cell line)
Loading amount 100000 cells
Specification MCF10A- Breast cell line
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Submitted May 24 2012

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Breast)
Specification Breast
Fixative Methanol
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 10%
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Verified customer

Submitted Dec 05 2011

Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
Loading amount 20 µg
Specification Mouse embryonic fibroblasts
Gel Running Conditions Reduced Denaturing (10% Tris-Gly gel)
Blocking step LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
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Submitted Nov 23 2011

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (Ntera2)
Total protein in input 20 µg
Specification Ntera2
Immuno-precipitation step Protein A
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Abcam user community

Verified customer

Submitted Aug 10 2011

Application Western blot
Sample Human Cell lysate - whole cell (Ntera2 cells)
Loading amount 20 µg
Specification Ntera2 cells
Gel Running Conditions Reduced Denaturing (12.5%)
Blocking step Milk as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Verified customer

Submitted Aug 10 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"