Overview

  • Product name
    Anti-Histone H2A (phospho S129) antibody
    See all Histone H2A primary antibodies
  • Description
    Rabbit polyclonal to Histone H2A (phospho S129)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ChIP, ELISA, PepArrmore details
  • Species reactivity
    Reacts with: Saccharomyces cerevisiae, Schizosaccharomyces pombe
    Does not react with: Human
  • Immunogen

    Synthetic peptide corresponding to Saccharomyces cerevisiae Histone H2A aa 100 to the C-terminus (phospho S129) conjugated to Bovine Serum Albumin (BSA).
    Database link: P04911

  • Positive control
    • MMS treated S. cerevisiae (0.1% for 30 minutes).

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab15083 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
WB 1/500 - 1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
ChIP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
PepArr Use a concentration of 0.2 - 0.02 µg/ml.

Target

  • Function
    Core component of nucleosome. 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.
  • Sequence similarities
    Belongs to the histone H2A family.
  • Post-translational
    modifications
    The chromatin-associated form is phosphorylated on Thr-121 during mitosis.
    Deiminated on Arg-4 in granulocytes upon calcium entry.
    Monoubiquitination of Lys-120 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression and participates in X chromosome inactivation of female mammals. It is involved in the initiation of both imprinted and random X inactivation. Ubiquitinated H2A is enriched in inactive X chromosome chromatin. Ubiquitination of H2A functions downstream of methylation of 'Lys-27' of histone H3. Monoubiquitination of Lys-120 by RNF2/RING2 can also be induced by ultraviolet and may be involved in DNA repair. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'Lys-63' linkage of ubiquitin moieties by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage. Monoubiquitination and ionizing radiation-induced 'Lys-63'-linked ubiquitination are distinct events.
    Phosphorylation on Ser-2 is enhanced during mitosis. Phosphorylation on Ser-2 by RPS6KA5/MSK1 directly represses transcription. Acetylation of H3 inhibits Ser-2 phosphorylation by RPS6KA5/MSK1.
    Symmetric dimethylation on Arg-4 by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
    • Alternative names
      • H2a 615 antibody
      • H2A antibody
      • H2A GL101 antibody
      • H2A histone family member A antibody
      • H2A.1 antibody
      • H2A.2 antibody
      • H2A/a antibody
      • H2A/m antibody
      • H2A/O antibody
      • H2A/q antibody
      • H2A1B_HUMAN antibody
      • H2AFA antibody
      • H2AFE antibody
      • H2AFL antibody
      • H2AFM antibody
      • H2AFO antibody
      • H2AFQ antibody
      • HIST1H2AE antibody
      • HIST1H2AJ antibody
      • HIST2H2AA antibody
      • HIST2H2AA3 antibody
      • HIST2H2AB antibody
      • HIST2H2AC antibody
      • Histone 1 H2ae antibody
      • Histone 2 H2aa3 antibody
      • Histone 2 H2ab antibody
      • Histone 2 H2ac antibody
      • Histone H2A type 1 B antibody
      • Histone H2A type 1 C antibody
      • Histone H2A type 1 E antibody
      • Histone H2A type 1 J antibody
      • Histone H2A type 1-B/E antibody
      • Histone H2A.2 antibody
      • Histone H2A/a antibody
      • Histone H2A/m antibody
      • MGC74460 antibody
      see all

    Images

    • All lanes : Anti-Histone H2A (phospho S129) antibody (ab15083) at 1/500 dilution

      Lane 1 : S.cerevisiae yeast extract (SCYE) + control peptide with Human Histone H2A peptide (ab19751)
      Lane 2 : S.cerevisiae yeast extract with 0.2 % Methyl methanesulfonate (1 hour) with Human Histone H2A peptide (ab19751)
      Lane 3 : SCYE + phospho peptide with Histone H2A peptide - phospho S129 (ab19828)
      Lane 4 : SCYE_M + phospho peptide with Histone H2A peptide - phospho S129 (ab19828)

      Lysates/proteins at 10 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 14 kDa



      The blots were produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membranes were then blocked for an hour. 1 microgram per mL of control- (ab19751, lane 1 and 2) or phospho-peptides (ab19828, lane 3 and 4) were added to the primary antibody ab15083 (rabbit anti-Histone H2A (phospho S129) antibody; diluted 1:500) and loading control ab125247 (mouse anti-GAPDH antibody; diluted 1:20000) and the membranes were incubated with peptide/antibody mixture for 24 hours at 4°C. Antibody binding was detected using infrared (IR)-labelled goat anti-rabbit (green) and  IR-labelled goat anti-mouse (red; insert below) antibodies, diluted 1:20,000, for 1 hour at room temperature before imaging.

    • Serially diluted ab15083 was bound to immobilised phospho (ab19828) - or control (ab19751) peptides (1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 50000 times) and signal was developed with TMB substrate.

    • Anti-Histone H2A (phospho S129) antibody (ab15083) at 1 µg/ml + S.cerevisiae (Y190) Whole Cell Lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 14 kDa
      Additional bands at: 17 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 90 seconds
    • All batches of ab15083 are tested in Peptide Array against peptides to different Histone H2A modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H2A - phospho S129 (ab19828), indicating that this antibody specifically recognises the Histone H2A - phospho S129 modification.

      ab19828 - Histone H2A - phospho S129

      ab19751 - Histone H2A - unmodified

    References

    This product has been referenced in:
    • Peng XP  et al. Acute Smc5/6 depletion reveals its primary role in rDNA replication by restraining recombination at fork pausing sites. PLoS Genet 14:e1007129 (2018). Read more (PubMed: 29360860) »
    • Sein H  et al. Rpb9-deficient cells are defective in DNA damage response and require histone H3 acetylation for survival. Sci Rep 8:2949 (2018). Read more (PubMed: 29440683) »
    See all 61 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U2OS cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    20 µg
    Treatment
    10 uM CPT for 1hr
    Specification
    U2OS cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Dec 20 2016

    Question
    Answer

    Yes, this antibody cross-reacts with Schizosaccharomyces pombe. You can find some references here:

    Carneiro T et al. Telomeres avoid end detection by severing the checkpoint signal transduction pathway. Nature 467:228-32 (2010). ChIP; Schizosaccharomyces pombe.PubMed: 20829797
    Wang Y et al. Regulation of Set9-mediated H4K20 methylation by a PWWP domain protein. Mol Cell 33:428-37 (2009). WB; Schizosaccharomyces pombe. PubMed: 19250904



    I have since updated the online datasheet to reflect this.

    Read More

    Answer

    >We only have full size vials of our products, so unfortunately we do not have a sample to send out. The antibody is fully guaranteed to work in Western blot and ChIP with S. cerevisiae samples for up to 612months after purchase. If you are interested in using the antibody with a species or application not listed on the datasheet, please let me know and I will send some information about our testing discount program.

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Saccharomyces cerevisiae Cell lysate - whole cell (BY4741 wild-type cells)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    2e+007 cells
    Treatment
    0, 1 and 10 µM nitrogen mustard for 2h
    Specification
    BY4741 wild-type cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Apr 18 2012

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

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