概述

  • 产品名称Anti-RNF20抗体
    参阅全部 RNF20 一抗
  • 描述
    兔多克隆抗体to RNF20
  • 经测试应用适用于: WB, ICC/IF, IHC-P, IPmore details
  • 种属反应性
    与反应: Chicken, Human
    预测可用于: Mouse
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human RNF20.

    (Peptide available as ab32628.)

  • 阳性对照
    • 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.

性能

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应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能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.
  • 组织特异性Expressed in the normal brain and also in malignant gliomas (at protein level).
  • 通路Protein modification; protein ubiquitination.
  • 序列相似性Belongs to the BRE1 family.
    Contains 1 RING-type zinc finger.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-RNF20 antibody 图像

  • 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 : A431 whole cell lysate (ab7909)
    Lane 4 : HEK293 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

Anti-RNF20 antibody (ab32629)参考文献

This product has been referenced in:
  • Long L  et al. The U4/U6 recycling factor SART3 has histone chaperone activity and associates with USP15 to regulate H2B deubiquitination. J Biol Chem 289:8916-30 (2014). WB . Read more (PubMed: 24526689) »
  • Klement K  et al. Opposing ISWI- and CHD-class chromatin remodeling activities orchestrate heterochromatic DNA repair. J Cell Biol 207:717-33 (2014). Read more (PubMed: 25533843) »

See all 6 Publications for this product

Product Wall

Abreviews
Application ChIP
Sample Human Cell lysate - whole cell (breast)
Specification breast
Detection step Real-time PCR
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
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Abcam user community

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提交于 Aug 21 2015

Abreviews
Application ChIP
Sample Mouse Cell lysate - whole cell (Colon)
Specification Colon
Detection step Real-time PCR
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
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Abcam user community

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提交于 Aug 21 2015

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

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提交于 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

提交于 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

提交于 Dec 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 Nov 28 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Abcam user community

Verified customer

提交于 May 24 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Breast)
Specification Breast
Fixative Methanol
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 10%
Username

Abcam user community

Verified customer

提交于 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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提交于 Nov 23 2011

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