製品の概要

  • 製品名
    Anti-CHD4 antibody [3F2/4] - ChIP Grade
    CHD4 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [3F2/4] to CHD4 - ChIP Grade
  • アプリケーション
    適用あり: ICC/IF, ChIP, ELISA, WB, IP, IHC-P, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Synthetic peptide (N- and C- terminal peptides were used for immunisation: 1. ASGLGSPSPCSAGSEEEDM & 2. CSRLANRAPEPPPQQVAQQQ) - which peptide the antibody recognises has not been tested.

  • ポジティブ・コントロール
    • WB: Human kidney tissue lysate, Human placenta tissue lysate, Human skin tissue lysate, HeLa (Human epithelial carcinoma cell line) WCL. IHC: human breast carcinoma tissue
  • 特記事項

    This antibody clone is manufactured by Abcam.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab70469 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
ICC/IF Use a concentration of 10 µg/ml.
ChIP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 238 kDa (predicted molecular weight: 218 kDa).
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能
    Component of the histone deacetylase NuRD complex which participates in the remodeling of chromatin by deacetylating histones.
  • 配列類似性
    Belongs to the SNF2/RAD54 helicase family.
    Contains 2 chromo domains.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
    Contains 2 PHD-type zinc fingers.
  • 翻訳後修飾
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 細胞内局在
    Nucleus. Cytoplasm > cytoskeleton > centrosome. Associates with centrosomes in interphase.
  • Information by UniProt
  • 参照データベース
  • 別名
    • ATP dependent helicase CHD4 antibody
    • ATP-dependent helicase CHD4 antibody
    • CHD 4 antibody
    • CHD-4 antibody
    • CHD4 antibody
    • CHD4_HUMAN antibody
    • Chromodomain helicase DNA binding protein 4 antibody
    • Chromodomain-helicase-DNA-binding protein 4 antibody
    • Mi 2 autoantigen 218 kDa protein antibody
    • Mi 2b antibody
    • Mi-2 autoantigen 218 kDa protein antibody
    • Mi2 beta antibody
    • Mi2-beta antibody
    see all

画像

  • CHD4 immunoprecipitation from murine brain tissue lysate using ab70469. 5000 µg of cell lysate was incubated with primary antibody (undiluted) and matrix (protein G) and incubated for 14 hours.

    An HRP-conjugated rabbit anti-CHD4 polyclonal (1/1000) was used to detect CHD4 immunoprecipitation.

    See Abreview

  • IHC image of CHD4 staining in human breast carcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ 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 ab70469, 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.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunocytochemical analysis of blastocysts from Wildtype and CHD4 -/- (Knockout) mice, labeling CHD4 with ab70469. Embryos were fixed in 2.5% paraformaldehyde, permeablised with 0.25% Triton X-100 and 3% polyvinlypyrrolidone in PBS, then blocked in 10% foetal calf serum and 0.1 Triton X-100 in PBS. Immunostaining with ab70469 (diluted 1/10,000) was performed overnight at 4°C. Staining with secondary antibody was for 1 hour at RT. DAPI used for nuclear staining (blue).

  • All lanes : Anti-CHD4 antibody [3F2/4] - ChIP Grade (ab70469) at 5 µg/ml

    Lane 1 : Human kidney tissue lysate - total protein (ab30203)
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Human placenta tissue lysate - total protein (ab29745)
    Lane 4 : Human skin tissue lysate - total protein (ab30166)
    Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 218 kDa
    Additional bands at : 238 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes

参考文献

This product has been referenced in:

See all 16 Publications for this product

レビューと Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (NIH-3T3 cells)
Permeabilization
Yes - tween-20
Specification
NIH-3T3 cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

投稿 Aug 31 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mouse urinary bladder)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 citric acid
Permeabilization
Yes - tween-20
Specification
mouse urinary bladder
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
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Abcam user community

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投稿 Jun 24 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Mouse Cell (MEL)
Specification
MEL
Permeabilization
No
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Dec 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293T cell)
Loading amount
30 µg
Specification
293T cell
Gel Running Conditions
Reduced Denaturing (7.5% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Min Young Kim

Verified customer

投稿 Dec 31 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Tissue lysate - whole (brain)
Total protein in input
500 µg
Specification
brain
Immuno-precipitation step
Protein G
Username

Dr. Catia Andreassi

Verified customer

投稿 Jul 16 2012

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

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