製品の概要

  • 製品名
  • 製品の詳細
    Mouse monoclonal to RNF168
  • アプリケーション
    適用あり: WB, ICC/IF, Flow Cyt, IHC-Pmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Recombinant fragment, corresponding to amino acids 462-572 of Human RNF168

  • 特記事項

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab220324 as a replacement.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab58063 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 - 5 µg/ml. Predicted molecular weight: 65 kDa.
ICC/IF Use at an assay dependent concentration. PubMed: 20725033
Flow Cyt Use 1µg for 106 cells. ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
IHC-P Use a concentration of 1.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ターゲット情報

  • 機能
    E3 ubiquitin-protein ligase required for accumulation of repair proteins to sites of DNA damage. Acts with UBE2N/UBC13 to amplify the RNF8-dependent histone ubiquitination. Recruited to sites of DNA damage at double-strand breaks (DSBs) by binding to ubiquitinated histone H2A and ubiquitinates histone H2A and H2AX, leading to amplify the RNF8-dependent H2A ubiquitination and promoting the formation of 'Lys-63'-linked ubiquitin conjugates. This leads to concentrate ubiquitinated histones H2A and H2AX at DNA lesions to the threshold required for recruitment of TP53BP1 and BRCA1.
  • パスウェイ
    Protein modification; protein ubiquitination.
  • 関連疾患
    Defects in RNF168 are the cause of Riddle syndrome (RIDDLES) [MIM:611943]. Riddle syndrome is characterized by increased radiosensitivity, immunodeficiency, mild motor control and learning difficulties, facial dysmorphism, and short stature. Defects are probably due to impaired localization of TP53BP1 and BRCA1 at DNA lesions.
  • 配列類似性
    Belongs to the RNF168 family.
    Contains 1 RING-type zinc finger.
  • ドメイン
    The MIU motifs (motif interacting with ubiquitin) mediate the interaction with ubiquitin and the localization at sites of DNA damage.
  • 細胞内局在
    Nucleus. Localizes to sites of DNA damage.
  • Information by UniProt
  • 参照データベース
  • 別名
    • E3 ubiquitin protein ligase RNF168 antibody
    • E3 ubiquitin-protein ligase RNF168 antibody
    • FLJ35794 antibody
    • FLJ39749 antibody
    • hRNF168 antibody
    • RING finger protein 168 antibody
    • RN168_HUMAN antibody
    • RNF 168 antibody
    • Rnf168 antibody
    see all

画像

  • Anti-RNF168 antibody (ab58063) at 1 µg/ml (HepG2 cell lysate)

    Predicted band size : 65 kDa
  • Immunohistochemical analysis formalin/PFA-fixed paraffin-enbedded Human prostate cancer tissue labelling RNF168 with ab58063 at 1.5μg/ml.

  • Immunofluorescence analysis of HepG2 cells labelling RNF168 with ab58063 at 10μg/ml.

  • Overlay histogram showing HepG2 cells stained with ab58063 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab58063, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min) /permeabilized in 0.1% PBS-Tween used under the same conditions.

参考文献

This product has been referenced in:
  • Silva BA  et al. DNA damage to a single chromosome end delays anaphase onset. J Biol Chem 289:22771-84 (2014). Read more (PubMed: 24982423) »
  • Zhu B  et al. K63-linked ubiquitination of FANCG is required for its association with the Rap80-BRCA1 complex to modulate homologous recombination repair of DNA interstand crosslinks. Oncogene N/A:N/A (2014). Read more (PubMed: 25132264) »

See all 4 Publications for this product

レビューと Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Treatment
cells transfected with RNF168 plasmids
Specification
U2OS
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Aug 31 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Permeabilization
Yes - 0.5% Triton X-100
Specification
U2OS
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Oct 26 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
50 µg
Specification
U2OS cells
Username

Abcam user community

Verified customer

投稿 Sep 28 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (U2OS cells)
Specification
U2OS cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Jul 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (U2OS)
Loading amount
20 µg
Specification
U2OS
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Dr. Andrew Cobb

Verified customer

投稿 Dec 20 2012

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

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