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, IHC-P, 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 (KLH).
(Peptide available as
- HeLa nuclear extract
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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
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).|
|IHC-P||1/5000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|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.
Image courtesy of Human Protein Atlas
ab15653 staining histone H2A in human male kidney, showing a distinct and strong nuclear staining pattern in tubuli and glomeruli. Paraffin embedded human skin tissue was incubated with ab15653 (1/5000 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6..
ab15653 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
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).
This product has been referenced in:
- 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 . Read more (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). Read more (PubMed: 28536406) »