製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to RNF20
  • アプリケーション
    適用あり: WB, ICC/IF, IHC-P, IPmore details
  • 種交差性
    交差種: Chicken, Human
    交差が予測される動物種: Mouse
  • 免疫原

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

    (Peptide available as ab32628.)

  • ポジティブ・コントロール
    • 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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    Expressed in the normal brain and also in malignant gliomas (at protein level).
  • パスウェイ
    Protein modification; protein ubiquitination.
  • 配列類似性
    Belongs to the BRE1 family.
    Contains 1 RING-type zinc finger.
  • 細胞内局在
    Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • 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

参考文献

This product has been referenced in:
  • Duan Y  et al. Ubiquitin ligase RNF20/40 facilitates spindle assembly and promotes breast carcinogenesis through stabilizing motor protein Eg5. Nat Commun 7:12648 (2016). Read more (PubMed: 27557628) »
  • Tamayo AG  et al. Histone monoubiquitination by Clock-Bmal1 complex marks Per1 and Per2 genes for circadian feedback. Nat Struct Mol Biol 22:759-66 (2015). WB ; Mouse . Read more (PubMed: 26323038) »

See all 10 Publications for this product

レビューと Q&A

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)
Username

Abcam user community

Verified customer

投稿 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)
Username

Abcam user community

Verified customer

投稿 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
Username

Abcam user community

Verified customer

投稿 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
Username

Dr. Abigail Buchwalter

Verified customer

投稿 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

投稿 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

投稿 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%
Username

Abcam user community

Verified customer

投稿 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

投稿 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
Username

Abcam user community

Verified customer

投稿 Nov 23 2011

1-10 of 20 Abreviews or Q&A

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

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