Recombinant
RabMAb

Anti-Histone H2A antibody [EPR17470] - ChIP Grade (ab177308)

Overview

  • Product name
    Anti-Histone H2A antibody [EPR17470] - ChIP Grade
    See all Histone H2A primary antibodies
  • Description
    Rabbit monoclonal [EPR17470] to Histone H2A - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: PepArr, ChIP, ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Histone H2A aa 50 to the C-terminus. The exact sequence is proprietary.
    Database link: P04908

  • Positive control
    • WB: HeLa cell lysate and mouse heart, brain, kidney and spleen lysates. IHC-P: Human colon, mouse colon and rat colon tissues. ICC/IF: HeLa cells. ChIP: HeLa cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR17470
  • Isotype
    IgG

Applications

Our Abpromise guarantee covers the use of ab177308 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
PepArr Use at an assay dependent concentration.
ChIP Use 2 µg for 25 µg of chromatin.
ICC/IF 1/500.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).

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

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H2A with ab177308 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing nuclear staining on HeLa cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    1. ab177308 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

     

  • Anti-Histone H2A antibody [EPR17470] - ChIP Grade (ab177308) at 1/5000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 14 kDa
    Observed band size: 14 kDa


    Exposure time: 30 seconds


    5% NFDM/TBST: Blocking and diluting buffer.

  • All lanes : Anti-Histone H2A antibody [EPR17470] - ChIP Grade (ab177308) at 1/1000 dilution

    Lane 1 : mouse brain lysate
    Lane 2 : mouse heart lysate
    Lane 3 : mouse kidney lysate
    Lane 4 : mouse spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 14 kDa
    Observed band size: 14 kDa


    Exposure time: 1 second


    5% NFDM/TBST: Blocking and diluting buffer.

  • 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 ab177308 (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.

  • ab177308 was tested in Peptide Array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
    Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
    The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H2A using ab177308 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstained with Hematoxylin. Nucleus staining on epithelial cells of Human colon tissue was observed.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H2A using ab177308 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstained with Hematoxylin. Nucleus staining on epithelial cells of mouse colon tissue was observed.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution..

  • Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H2A using ab177308 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstained with Hematoxylin. Nucleus staining on epithelial cells of rat colon tissue was observed.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

References

This product has been referenced in:
  • Zhang ZH  et al. Dynamics of histone H2A, H4 and HS1ph during spermatogenesis with a focus on chromatin condensation and maturity of spermatozoa. Sci Rep 6:25089 (2016). WB . Read more (PubMed: 27121047) »
  • Kafer GR  et al. 5-Hydroxymethylcytosine Marks Sites of DNA Damage and Promotes Genome Stability. Cell Rep 14:1283-1292 (2016). Read more (PubMed: 26854228) »
See all 2 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Vascular smooth muscle cells
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Jun 25 2018

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

Sign up