Key features and details
- Rabbit polyclonal to Histone H2A - ChIP Grade
- Suitable for: WB, ICC/IF, ChIP
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Histone H2A antibody - ChIP Grade
See all Histone H2A primary antibodies
DescriptionRabbit polyclonal to Histone H2A - ChIP Grade
Specificityab15653 was generated using an immunogen specific to the C-terminal of H2A2B (SwissProt Q8IUE6). Interestingly mass spec analysis on immunoprecipitated material using ab15653 indicates that the antibody recognises all isotypes of H2A.
Tested applicationsSuitable for: WB, ICC/IF, ChIPmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse
Synthetic peptide corresponding to Human Histone H2A aa 100 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available as
- HeLa nuclear extract
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Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
ChIP Related Products
Our Abpromise guarantee covers the use of ab15653 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.5 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
|ChIP||Use 2-25 µg for µg of chromatin.|
FunctionCore 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 similaritiesBelongs to the histone H2A family.
modificationsThe 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 localizationNucleus. Chromosome.
- Information by UniProt
- H2a 615 antibody
- H2A antibody
- H2A GL101 antibody
All lanes : Anti-Histone H2A antibody - ChIP Grade (ab15653) at 0.5 µg/ml
Lane 1 : HeLa nuclear extract
Lane 2 : HeLa nuclear extract with Human Histone H2A peptide (ab15660) at 1 µg/ml
Predicted band size: 14 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
ab15653 recognises H2A in HeLa nuclear extracts (lane1), which is successfully blocked using the immunizing peptide (lane2).
Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab15653 (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). Primers and probes are located in the first kb of the transcribed region.
ICC/IF image of ab15653 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab15653, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab15653 has been referenced in 7 publications.
- Sun H et al. miR-4429 sensitized cervical cancer cells to irradiation by targeting RAD51. J Cell Physiol 235:185-193 (2020). PubMed: 31190335
- Kim J et al. Replication Stress Shapes a Protective Chromatin Environment across Fragile Genomic Regions. Mol Cell 69:36-47.e7 (2018). PubMed: 29249653
- Jullien D et al. Chromatibody, a novel non-invasive molecular tool to explore and manipulate chromatin in living cells. J Cell Sci 129:2673-83 (2016). WB ; Human . PubMed: 27206857
- Lee YH et al. Regulation of DNA Damage Response by Estrogen Receptor ß-Mediated Inhibition of Breast Cancer Associated Gene 2. Biomedicines 3:182-200 (2015). PubMed: 28536406
- Loza-Muller L et al. Fibrillarin methylates H2A in RNA polymerase I trans-active promoters in Brassica oleracea. Front Plant Sci 6:976 (2015). Mouse . PubMed: 26594224
- Reitsma JM et al. Human cytomegalovirus IE1 protein disrupts interleukin-6 signaling by sequestering STAT3 in the nucleus. J Virol 87:10763-76 (2013). PubMed: 23903834
- Burd CJ et al. Analysis of chromatin dynamics during glucocorticoid receptor activation. Mol Cell Biol 32:1805-17 (2012). ChIP . PubMed: 22451486