Anti-Histone H2A antibody - ChIP Grade (ab15653)
Key features and details
- Rabbit polyclonal to Histone H2A - ChIP Grade
- Suitable for: WB, ICC/IF, ChIP
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Histone H2A antibody - ChIP Grade
See all Histone H2A primary antibodies -
Description
Rabbit polyclonal to Histone H2A - ChIP Grade -
Host species
Rabbit -
Specificity
ab15653 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 applications
Suitable for: WB, ICC/IF, ChIPmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse
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Immunogen
Synthetic peptide corresponding to Human Histone H2A aa 100 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab15660) -
Positive control
- HeLa nuclear extract
Properties
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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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
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.
Application Abreviews Notes 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. Target
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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
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 localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 3012 Human
- Entrez Gene: 317772 Human
- Entrez Gene: 8335 Human
- Entrez Gene: 8338 Human
- Entrez Gene: 319166 Mouse
- Omim: 142720 Human
- Omim: 602786 Human
- Omim: 602797 Human
see all -
Alternative names
- H2a 615 antibody
- H2A antibody
- H2A GL101 antibody
see all
Images
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Western blot - Anti-Histone H2A antibody - ChIP Grade (ab15653)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). -
ChIP - Anti-Histone H2A antibody - ChIP Grade (ab15653)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.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A antibody - ChIP Grade (ab15653)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).
Protocols
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.
Datasheets and documents
References (7)
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
Images
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Western blot - Anti-Histone H2A antibody - ChIP Grade (ab15653)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). -
ChIP - Anti-Histone H2A antibody - ChIP Grade (ab15653)
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.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A antibody - ChIP Grade (ab15653)
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).
