Anti-H2A.Z (acetyl K4 + K7 + K11) 抗体 - ChIP Grade (ab18262)

製品の概要

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab18262 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
ChIP Use at an assay dependent dilution.
ICC/IF Use at an assay dependent dilution.
WB Use at an assay dependent dilution. Detects a band of approximately 14 kDa (predicted molecular weight: 13 kDa).

ターゲット情報

  • 機能Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. May be involved in the formation of constitutive heterochromatin. May be required for chromosome segregation during cell division.
  • 配列類似性Belongs to the histone H2A family.
  • 翻訳後修飾Monoubiquitination of Lys-122 gives a specific tag for epigenetic transcriptional repression.
    Acetylated on Lys-5, Lys-8 and Lys-12 during interphase. Acetylation disappears at mitosis.
    Not phosphorylated.
  • 細胞内局在Nucleus. Chromosome.
  • Information by UniProt
  • 参照データベース
  • 別名
    • H2A/z antibody
    • H2afz antibody
    • H2AZ antibody
    • H2AZ_HUMAN antibody
    • Histone H2A.Z antibody
    see all

Anti-H2A.Z (acetyl K4 + K7 + K11) antibody - ChIP Grade 画像

  • ChIP analysis of Acetyl-H2A.Z at the p21 transcription start site with or without the HDAC inhibitor TSA (50ng/ml). Chromatin was prepared from MDA-MB231 cells. Cells were fixed with formaldehyde for 10min. Samples were sonicated to generate DNA fragments between 500 and 700 bp. The ChIP was performed with 50µg of chromatin, fragments were immunoprecipitated using 2µg of antibody ab18262, and an HA antibody was used as control. The precipitated DNA was amplified by real-time PCR, with primer sets designed to amplify the promoter and the coding region of the p21 gene.

     

     

  • Anti-H2A.Z (acetyl K4 + K7 + K11) antibody - ChIP Grade (ab18262) at 1/1000 dilution + HeLa whole cell lysate at 10 µg

    Secondary
    HRP-conjugated donkey anti-sheep IgG polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 13 kDa
    Observed band size : 13 kDa


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

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  • ab18262 staining Histone H2A.Z (acetyl K4 + K7 + K11) in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 3% BSA for 1 hour at 22°C. Samples were incubated with primary antibody (1/500 in PBS + 1% BSA) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated donkey anti-sheep IgG polyclonal (1/1000) was used as the secondary antibody.

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  • All lanes : Anti-H2A.Z (acetyl K4 + K7 + K11) antibody - ChIP Grade (ab18262)

    Lane 1 : Butyrate-treated HeLa histones
    Lane 2 : 15-day embryo chicken erythrocyte histones
    Lane 3 : Recombinant H2A.Z
    Lane 4 : Butyrate-treated HeLa histones
    Lane 5 : 15-day embryo chicken erythrocyte histones
    Lane 6 : Recombinant H2A.Z


    Predicted band size : 13 kDa
    Observed band size : 13 kDa

    This image is courtesy of Prof Colyn Crane-Robinson

  • All lanes : Anti-H2A.Z (acetyl K4 + K7 + K11) antibody - ChIP Grade (ab18262)

    Lane 1 : Butyrate-treated HeLa histones
    Lane 2 : Nucleosomes from 10-day chicken brain tissue. ChIP Input.
    Lane 3 : Nucleosomes from 10-day chicken brain tissue. ChIP Unbound.
    Lane 4 : Nucleosomes from 10-day chicken brain tissue. ChIP Bound.
    Lane 5 : Butyrate-treated HeLa histones
    Lane 6 : Nucleosomes from 10-day chicken brain tissue. ChIP Input.
    Lane 7 : Nucleosomes from 10-day chicken brain tissue. ChIP Unbound.
    Lane 8 : Nucleosomes from 10-day chicken brain tissue. ChIP Bound.


    Predicted band size : 13 kDa
    Observed band size : 13 kDa

    This image is courtesy of Prof Colyn Crane-Robinson

    Nucleosomes from 10-day chicken brain tissue were used in ChIP experiments. An acetic acid / urea / Triton gel was run and the corresponding western blot using ab18262 was used to compare proteins in the Bound chromatin fraction with those of the Input and Unbound fractions. In lane 8 there is a strong diffuse band corresponding to a hyper-acetylated H2A.Z.

Anti-H2A.Z (acetyl K4 + K7 + K11) antibody - ChIP Grade (ab18262) 使用論文

This product has been referenced in:
  • Kusch T  et al. Histone H3 lysine 4 trimethylation regulates cotranscriptional H2A variant exchange by Tip60 complexes to maximize gene expression. Proc Natl Acad Sci U S A 111:4850-5 (2014). Drosophila melanogaster . Read more (PubMed: 24639513) »
  • Menafra R  et al. Genome-Wide Binding of MBD2 Reveals Strong Preference for Highly Methylated Loci. PLoS One 9:e99603 (2014). CHIPseq ; Human . Read more (PubMed: 24927503) »

See all 15 Publications for this product

Product Wall

Application Western blot
Loading amount 0.5 µg
Gel Running Conditions Reduced Denaturing (Criterion TGX 4-20% (BioRad))
Sample Human Purified protein (Recombinant octamers & purified K562 cell histones)
Specification Recombinant octamers & purified K562 cell histones
Treatment TSA 50 ng/ml 4 hours
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Username

Dr. Ragnhild Eskeland

Verified customer

投稿 Jan 27 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.5% Triton X100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Sep 30 2014

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (12% SDS-PAGE)
Sample Human Cell lysate - whole cell (HeLa)
Specification HeLa
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Sep 25 2014

Regarding the specificity of ab18262, referencing Bruce et al, 2005, Nucleic Acids Research v33, p5633-5639, Fig  1. Although SDS gels/Westerns do not distinguish H2A from H2A.Z (Panels A & B), in AUT gels/Westerns they are readily distinguished (Panel...

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Thank you for your inquiry.

With only 41% homology between the immunogen and the yeast histone (see the attached pdf), it is unlikely that ab18262 would work on yeast.

I would like to recommend checking the Biocompare website which ha...

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I'm sorry about that. It might be that the system won't let you submit a review for an unpiblished antibody. I was not aware of this.

If you wouldn't mind, could you just share the information with me by filling in the relevant form attached ...

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It is always a pleasure to help.



Please do not hesitate let us know if you have ever any questions or there are other ways that Abcam may help you meet your research goals.

Thank you for contacting us.

This product has been designed to extract histones with their modifications intact. Therefore this should be very suitable for your needs.

I hope this information is helpful to you. Please do not h...

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Thank you for contacting Abcam and for your interest in our products.


Regarding theH2A.Z (acetyl K4 + K7 + K11) antibody ab18262, the number of bands can be indeed due to different acetylation states as we have found multiple bands in our...

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Thank you for your enquiry.

I can confirm that ab18262 regognizes Anti-Histone H2A.Z (acetyl K4 + K7 + K11) antibody - ChIP Grade.

It may well be that the label of the vial has the shortened form of this target name due to limited spa...

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