Anti-Histone H2A.Z antibody - ChIP Grade (ab97966)


  • Product name
    Anti-Histone H2A.Z antibody - ChIP Grade
    See all Histone H2A.Z primary antibodies
  • Description
    Rabbit polyclonal to Histone H2A.Z - ChIP Grade
  • Host species
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, ChIP, IPmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Schizosaccharomyces pombe
  • Immunogen

    Synthetic peptide corresponding to Human Histone H2A.Z aa 65-128.

  • Positive control
    • MOLT4 or Raji cell lysate. ICC/IF Hela cells IHC-P: Human normal colon FFPE tissue sections.



Our Abpromise guarantee covers the use of ab97966 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 0.5 µg/ml.
WB 1/500 - 1/3000. Predicted molecular weight: 14 kDa.
IHC-P Use a concentration of 0.1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ChIP Use 2 µg for 25 µg of chromatin.
IP 1/500 - 1/1000.


  • Function
    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.
  • Sequence similarities
    Belongs to the histone H2A family.
  • Post-translational
    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.
    Monomethylated on Lys-5 and Lys-8 by SETD6. SETD6 predominantly methylates Lys-8, lys-5 being a possible secondary site.
    Not phosphorylated.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H2A histone family member Z antibody
    • H2A.z antibody
    • H2A/z antibody
    • H2afz antibody
    • H2AZ antibody
    • H2AZ_HUMAN antibody
    • Histone H2A.Z antibody
    • MGC117173 antibody
    see all


  • All lanes : Anti-Histone H2A.Z antibody - ChIP Grade (ab97966) at 1/3000 dilution

    Lane 1 : MOLT4 whole cell lysate
    Lane 2 : Raji whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 14 kDa

    12% SDS PAGE
  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab97966 (blue), and 20µl 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.

  • IHC image of ab97966 staining Histone H3 (phospho T45) 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 ab97966, 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.

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

  • ab97966 staining Histone H2A.Z in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab97966 at 0.5μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 2 μg/ml (shown in green). AlexaFluor®350 WGA was used at a 1/200 dilution and incubated for 1h with the cells, to label plasma membranes (shown in blue). Nuclear DNA was labelled in red with 1.25 μM DRAQ5™ (ab108410), which was added to the secondary antibody mixture. A secondary only negative control is displayed, which indicates that the Histone H2A.Z staining observed is due to primary antibody specificity and not to unspecific binding of the secondary antibody to the cells.

  • ab97966 immunoprecipitating Histone H2A.Z protein in HeLa whole cell lysate/extract. Lane 1: 50 μg HeLa whole cell lysate/extract. Lane 2: Control with 2 μg of preimmune rabbit IgG. Lane 3: Immunoprecipitation of Histone H2A.Z protein by 2 μg of ab97966. The immunoprecipitated Histone H2A.Z protein was detected with ab97966 diluted at 1:1000. Anti-rabbit IgG was used as a secondary antibody.


ab97966 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Western blot
Saccharomyces cerevisiae Cell lysate - whole cell (WT and htz1? (Htz1 deletion ) cells)
Gel Running Conditions
Reduced Denaturing (gel 15%)
Loading amount
1e+007 cells
WT and htz1? (Htz1 deletion ) cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 30°C

Daisuke Takahashi

Verified customer

Submitted Jul 17 2018

Thank you very much for providing that information.

I am sorry that the antibody did not prove successful with your S. pombe samples. I do hope that you feel it was still worthwhile trying. As I mentioned in my previous email, please do go ah...

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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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Thank you for contacting me.

I am sorry that the Anti-Histone H2A.Z antibody (ab97966) has now been removed from the catalogue. I am sorry but I was not aware of this change when I suggested you test it. Let me reassure you that the antibody ...

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I am glad you found the information provided of help.

All you need to do in order to participate in the testing discount scheme is to purchase ab97966 as normal. Once you have used with yourS. pombe samples, please share the results observed ...

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Thank you for contacting us and sorry for the delay in getting back to you.

I have checked the sequence provided and compared it to the immunogen of all the anti-Histone H2A.Z antibodies we have, including the ones you suggested. Unfortunatel...

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