製品の概要

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

    Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Human CoREST.

    (Peptide available as ab25741.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in Hela and Jurkat whole cell and nuclear lysates

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab32631 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 70 kDa (predicted molecular weight: 70 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能
    Essential component of the BHC complex, a corepressor complex that represses transcription of neuron-specific genes in non-neuronal cells. The BHC complex is recruited at RE1/NRSE sites by REST and acts by deacetylating and demethylating specific sites on histones, thereby acting as a chromatin modifier. In the BHC complex, it serves as a molecular beacon for the recruitment of molecular machinery, including MeCP2 and SUV39H1, that imposes silencing across a chromosomal interval. Plays a central role in demethylation of Lys-4 of histone H3 by promoting demethylase activity of KDM1A on core histones and nucleosomal substrates. It also protects KDM1A from the proteasome.
  • 組織特異性
    Ubiquitously expressed.
  • 配列類似性
    Belongs to the CoREST family.
    Contains 1 ELM2 domain.
    Contains 2 SANT domains.
  • ドメイン
    The SANT domains may bridge the nucleosomal substrates and the demethylase KDM1A.
  • 翻訳後修飾
    Phosphorylated by HSV-1 protein kinases in case of infection.
  • 細胞内局在
    Nucleus. Upon infection by HSV-1, it is partially translocated into the cytoplasm in an HSV-1-dependent manner.
  • Information by UniProt
  • 参照データベース
  • 別名
    • COREST antibody
    • KIAA0071 antibody
    • Nr2b2 antibody
    • Nuclear receptor coregulator 1 antibody
    • Nuclear receptor subfamily 2 group B member 2 antibody
    • Protein CoREST antibody
    • RCOR antibody
    • Rcor1 antibody
    • RCOR1_HUMAN antibody
    • REST corepressor 1 antibody
    • Retinoic acid receptor RXR-beta antibody
    • Retinoid X receptor beta antibody
    • Rxrb antibody
    see all

画像

  • Lanes 1 - 2 : Anti-CoREST antibody (ab32631) at 1 µg/ml
    Lanes 3 - 4 : Anti-CoREST antibody (ab32631) at 1/1 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Nuclear Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 70 kDa
    Additional bands at : 15 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 10 minutes
  • PCR was performed on a SCNA1 promoter following CoRest chromatin immunoprecipition using ab32631 at a concentration of 3ug/ml from 293T cells.
    Lane 1 - ab24166
    Lane 2 - Beads only control
    Lane 3 - IgG only control
    Lane 4 - Input
  • CoREST was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to CoREST and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32631.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 65kDa: CoREST.
  • IHC image of CoREST staining in Human normal tonsil FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32631, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
  • ICC/IF image of ab32631 stained MCF7 cells. The cells were 10% 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 (ab32631, 5µg/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) HeLa, Hek293 and HepG2 cells at 5µg/ml.

参考文献

This product has been referenced in:
  • Ao R  et al. Altered microRNA-9 Expression Level is Directly Correlated with Pathogenesis of Nonalcoholic Fatty Liver Disease by Targeting Onecut2 and SIRT1. Med Sci Monit 22:3804-3819 (2016). WB ; Human . Read more (PubMed: 27756894) »
  • Harten SK  et al. The recently identified modifier of murine metastable epialleles, Rearranged L-Myc Fusion, is involved in maintaining epigenetic marks at CpG island shores and enhancers. BMC Biol 13:21 (2015). WB . Read more (PubMed: 25857663) »

See all 6 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (esophagus)
Permeabilization
Yes - 1% triton x 100
Specification
esophagus
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Formaldehyde
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投稿 Sep 08 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (liver cells)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
40 µg
Specification
liver cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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投稿 Sep 05 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Brain)
Specification
Brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Paraformaldehyde
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Verified customer

投稿 Aug 20 2015

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

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