Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RNF20 antibody (ab32629)

Key features and details

  • Rabbit polyclonal to RNF20
  • Suitable for: WB, ICC/IF
  • Reacts with: Chicken, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-RNF20 antibody
    See all RNF20 primary antibodies

  • Description

    Rabbit polyclonal to RNF20

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Chicken, Human
    Predicted to work with: Mouse

  • Immunogen

    Synthetic peptide corresponding to Human RNF20 aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab32628)

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

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 Abreviews Notes
WB (8)
Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 114 kDa).
ICC/IF (6)
Use a concentration of 1 µg/ml.
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 114 kDa).
ICC/IF
Use a concentration of 1 µg/ml.

Target

  • Function

    Component of the RNF20/40 E3 ubiquitin-protein ligase complex that mediates monoubiquitination of 'Lys-120' of histone H2B (H2BK120ub1). H2BK120ub1 gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation (H3K4me and H3K79me, respectively). It thereby plays a central role inb histone code and gene regulation. The RNF20/40 complex forms a H2B ubiquitin ligase complex in cooperation with the E2 enzyme UBE2A or UBE2B; reports about the cooperation with UBE2E1/UBCH are contradictory. Required for transcriptional activation of Hox genes. Recruited to the MDM2 promoter, probably by being recruited by p53/TP53, and thereby acts as a transcriptional coactivator. Mediates the polyubiquitination of isoform 2 of PA2G4 in cancer cells leading to its proteasome-mediated degradation.

  • Tissue specificity

    Expressed in the normal brain and also in malignant gliomas (at protein level).

  • Pathway

    Protein modification; protein ubiquitination.

  • Sequence similarities

    Belongs to the BRE1 family.
    Contains 1 RING-type zinc finger.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • BRE1 A antibody
    • BRE1 antibody
    • BRE1 E3 ubiquitin ligase homolog antibody
    • BRE1-A antibody
    • BRE1A antibody
    • BRE1A_HUMAN 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
    • Rnf20 antibody
    see all

Images

  • Western blot - Anti-RNF20 antibody (ab32629)
    Western blot - Anti-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 whole cell lysate (ab7899)
    Lane 3 : A-431 whole cell lysate (ab7909)
    Lane 4 : HEK-293 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
    Lane 1 : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
    Lanes 2-6 : 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?



        
  • Immunocytochemistry/ Immunofluorescence - Anti-RNF20 antibody (ab32629)
    Immunocytochemistry/ Immunofluorescence - Anti-RNF20 antibody (ab32629)This image is courtesy of an Abreview submitted by Dr Kirk McManus
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-RNF20 antibody (ab32629)
    Immunocytochemistry/ Immunofluorescence - Anti-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)
    Western blot - Anti-RNF20 antibody (ab32629)Image courtesy of an anonymous Abreview.
    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
    All lanes : 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

    See Abreview

  • Western blot - Anti-RNF20 antibody (ab32629)
    Western blot - Anti-RNF20 antibody (ab32629)
    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 (17)

Publishing research using ab32629? Please let us know so that we can cite the reference in this datasheet.

ab32629 has been referenced in 17 publications.

  • Suraweera A  et al. COMMD4 functions with the histone H2A-H2B dimer for the timely repair of DNA double-strand breaks. Commun Biol 4:484 (2021). PubMed: 33875784
  • Mani C  et al. The multifunctional protein PACS-1 is required for HDAC2- and HDAC3-dependent chromatin maturation and genomic stability. Oncogene 39:2583-2596 (2020). PubMed: 31988453
  • Fiore D  et al. N6-Isopentenyladenosine Inhibits Colorectal Cancer and Improves Sensitivity to 5-Fluorouracil-Targeting FBXW7 Tumor Suppressor. Cancers (Basel) 11:N/A (2019). PubMed: 31569395
  • Wang JH  et al. Activation of the mammalian target of rapamycin signaling pathway underlies a novel inhibitory role of ring finger protein 182 in ventricular remodeling after myocardial ischemia-reperfusion injury. J Cell Biochem N/A:N/A (2018). PubMed: 30450663
  • Shah VJ & Maddika S CRL7SMU1 E3 ligase complex-driven H2B ubiquitylation functions in sister chromatid cohesion by regulating SMC1 expression. J Cell Sci 131:N/A (2018). PubMed: 29507117
View all Publications for this product

Images

  • Western blot - Anti-RNF20 antibody (ab32629)
    Western blot - Anti-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 whole cell lysate (ab7899)
    Lane 3 : A-431 whole cell lysate (ab7909)
    Lane 4 : HEK-293 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
    Lane 1 : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
    Lanes 2-6 : 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?



        
  • Immunocytochemistry/ Immunofluorescence - Anti-RNF20 antibody (ab32629)
    Immunocytochemistry/ Immunofluorescence - Anti-RNF20 antibody (ab32629) This image is courtesy of an Abreview submitted by Dr Kirk McManus
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-RNF20 antibody (ab32629)
    Immunocytochemistry/ Immunofluorescence - Anti-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)
    Western blot - Anti-RNF20 antibody (ab32629) Image courtesy of an anonymous Abreview.
    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
    All lanes : 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

    See Abreview

  • Western blot - Anti-RNF20 antibody (ab32629)
    Western blot - Anti-RNF20 antibody (ab32629)
    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

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