Overview

Properties

Applications

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.

Application Abreviews Notes
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).

Target

  • 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
    modifications
    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.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H2A/z antibody
    • H2afz antibody
    • H2AZ antibody
    • H2AZ_HUMAN antibody
    • Histone H2A.Z antibody
    see all

Images

  • 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 (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

    See Abreview

  • 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.

    See Abreview

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

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

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

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

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

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



    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.

References

This product has been referenced in:
  • Zeng Z  et al. Poly (ADP-ribose) glycohydrolase silencing-mediated maintenance of H2A and downregulation of H2AK9me protect human bronchial epithelial cells from benzo(a)pyrene-induced carcinogenesis. Toxicol Lett 295:270-276 (2018). Read more (PubMed: 29981922) »
  • Dzida T  et al. Predicting stimulation-dependent enhancer-promoter interactions from ChIP-Seq time course data. PeerJ 5:e3742 (2017). Read more (PubMed: 28970965) »
See all 25 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

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

Dr. Ragnhild Eskeland

Verified customer

Submitted Jan 27 2015

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

Abcam user community

Verified customer

Submitted Sep 30 2014

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

Abcam user community

Verified customer

Submitted Sep 25 2014

Answer

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 (Panels C & D).  In the HeLa butyrate lanes (HB, where the acetylation of all the histones is high) the Western shows a pair of H2A.Z bands and no H2A band (despite the amount of H2A being much greater than that of H2A.Z).  In the Bound fraction from the ChIP, there is a strong signal from H2A.Z but none from H2A.
Thus, ab18262 is specific for H2A.Z and does not recognise H2A at all - as expected since the H2A.Z N-terminal peptide immunogen is not represented in H2A.

Read More

Answer

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 has an excellent antibody search facility that includes many suppliers. The link is http://www.biocompare.com.

Also, we are happy to be able to offer custom antibody services through Epitomics, an Abcam company. For more information about these services visit http://www.epitomics.com/services/
email: mailto:service@epitomics.com

I hope this information is nevertheless helpful to you. Please do not hesitate to contact me if you have any further questions in this regard.

Read More

Answer

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 to this email? This should only take 5 minutes. Could you also let me know how you would rate the antibody out of 5.

This information would only become accessible to others if we republish this antibody in the future.

Once the form has been returned to me, please go ahead and use the discount code I provided for you. Along with the discount code could you please quote "antibody unpublished, testing discount agreed with Karin". This means customer services know to speak to me once the order goes through so the discount will be added.

I look forward to receiving your reply.

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Question
Answer

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.

Read More

Answer

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 hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

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 own WB and attribute more bands to hyperacetylated Histone H2A.Z.

I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Read More

Answer

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 space ie. formatting.

Could you please confirm if the label has the Abcam new logo and the batch number printed?

If you need any further assistance in the future, please do not hesitate to contact me.

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1-10 of 15 Abreviews or Q&A

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

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