Anti-Histone H4 (acetyl K91) 抗体 - ChIP Grade (ab4627)

製品の概要

  • 製品名
    Anti-Histone H4 (acetyl K91) antibody - ChIP Grade
    Histone H4 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Histone H4 (acetyl K91) - ChIP Grade
  • アプリケーション
    適用あり: IHC-P, WB, CHIPseq, ICC/IF, IP, ChIPmore details
  • 種交差性
    交差種: Mouse, Cow, Saccharomyces cerevisiae
    交差が予測される動物種: Rat, Chicken, Human, Pig, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Duck
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Histone H4, acetylated at K91.

    (Peptide available as ab10112.)

  • ポジティブ・コントロール
    • Calf thymus histone prep. IHC-P: Human normal colon FFPE tissue sections.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab4627 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 11 kDa (predicted molecular weight: 12 kDa).Can be blocked with Human Histone H4 (acetyl K91) peptide (ab10112).
CHIPseq Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IP 1/75.
ChIP Use at an assay dependent concentration.

ターゲット情報

画像

  • ab4627 staining Histone H4 (acetyl K91) in mouse C2C12 muscle fibroblast cultured cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 5% serum for 20 minutes at 23°C. Samples were incubated with primary antibody (1/1000 in 3% BSA + 0.1% Triton X-100 in PBS) for 12 hours at 4°C. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/2000) was used as the secondary antibody.

    See Abreview

  • IHC image of ab4627 staining Histone H2A.X in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab4627, 1µg/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. No primary antibody was used in the negative control (shown on the inset).

    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.

  • Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The  ChIP was performed with 25 µg of chromatin, 2 µg of  ab4627 (blue), and 20 µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.



  • Predicted band size : 12 kDa

    Rabbit polyclonal to Histone H4 acetyl K91 (ab4627). All lanes contain 1ug of calf thymus Histone prep.

    Lane 1: ab4627 at 1/500
    Lane 2: ab4627 at 1/1000
    Lane 3: ab4627 at 1/500 + 2ug acetyl K91 blocking peptide (ab10112)
    Lane 4: ab4627 at 1/1000 + 2ug acetyl K91 blocking peptide (ab10112)

    Secondary antibody: Goat anti-rabbit (HRP) - ab6721 at 1/2000

    Predicted Molecular weight11KDa

参考文献

This product has been referenced in:
  • Yang X  et al. Histone acetyltransferase 1 promotes homologous recombination in DNA repair by facilitating histone turnover. J Biol Chem 288:18271-82 (2013). Human . Read more (PubMed: 23653357) »
  • Singh P  et al. Allele-specific H3K79 Di- versus trimethylation distinguishes opposite parental alleles at imprinted regions. Mol Cell Biol 30:2693-707 (2010). Read more (PubMed: 20351169) »

See all 9 Publications for this product

レビューと Q&A

Application
Immunocytochemistry
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Mouse Cultured Cells (C2C12 muscle fibroblast)
Specification
C2C12 muscle fibroblast
Permeabilization
Yes - 0.1% Triton X-100
Fixative
Paraformaldehyde
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投稿 Feb 25 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Caenorhabditis elegans Tissue lysate - whole (C. elegans larvae)
Specification
C. elegans larvae
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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投稿 Feb 21 2014

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (Fruit fly embryo)
Specification
Fruit fly embryo
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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投稿 Feb 21 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 37 minute(s) · Concentration: 3% · Temperature: 37°C
Sample
Mouse Cell (NIH3T3)
Specification
NIH3T3
Permeabilization
Yes - 0.25% ~0.5% Triton
Fixative
Formaldehyde
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投稿 Dec 20 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (BY4147 background)
Total protein in input
1e+006 cells
Specification
BY4147 background
Treatment
0.75% MMS (DNA damaging agent)
Immuno-precipitation step
Protein G
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投稿 Aug 09 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (S288c)
Loading amount
25 µg
Specification
S288c
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
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Mr. Harsh Ja

Verified customer

投稿 Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (S288c)
Specification
S288c
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Fromaldehyde
Detection step
Real-time PCR
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Mr. Harsh Ja

Verified customer

投稿 Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (W303 background)
Specification
W303 background
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 20 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step
Semiquantitative PCR
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投稿 May 31 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application
ChIP on chip
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (W303 background)
Specification
W303 background
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 20 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step
Microarray
Negative control
No antibody control
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投稿 May 31 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (Yeast BY4741 background)
Loading amount
10 µg
Specification
Yeast BY4741 background
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 May 26 2011

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

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