Anti-Histone H2A.X antibody (ab47503)
- Product nameAnti-Histone H2A.X antibodySee all Histone H2A.X primary antibodies ...
- DescriptionRabbit polyclonal to Histone H2A.X
- SpecificityThis antibody detects endogenous levels of total Histone H2A.X protein. ab47503 reacts with non-phosphorylated form of Histone H2A.X and not the phosphorylated form of the protein.
- Tested applicationsELISA, WB, IHC-P more details
- Species reactivityReacts with: Human
Predicted to work with: Mouse
A sinthetic non-phosphopeptide derived from human Histone H2A.X around the phosphorylation site of serine 140 (Q-A-SP-Q-E)
- Positive control
- Hela cell extracts; Human breast carcinoma tissue
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
- Concentration information loading...
- PurityImmunogen affinity purified
- Purification notesThe antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab47503 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||WB: 1/500 - 1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 15 kDa).|
|IHC-P||IHC-P: Use at an assay dependent dilution.|
- FunctionVariant histone H2A which replaces conventional H2A in a subset of nucleosomes. 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. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation.
- Sequence similaritiesBelongs to the histone H2A family.
- Developmental stageSynthesized in G1 as well as in S-phase.
- DomainThe [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family.
modificationsPhosphorylated on Ser-140 (to form gamma-H2AFX or H2AX139ph) in response to DNA double strand breaks (DSBs) generated by exogenous genotoxic agents and by stalled replication forks, and may also occur during meiotic recombination events and immunoglobulin class switching in lymphocytes. Phosphorylation can extend up to several thousand nucleosomes from the actual site of the DSB and may mark the surrounding chromatin for recruitment of proteins required for DNA damage signaling and repair. Widespread phosphorylation may also serve to amplify the damage signal or aid repair of persistent lesions. Phosphorylation of Ser-140 (H2AX139ph) in response to ionizing radiation is mediated by both ATM and PRKDC while defects in DNA replication induce Ser-140 phosphorylation (H2AX139ph) subsequent to activation of ATR and PRKDC. Dephosphorylation of Ser-140 by PP2A is required for DNA DSB repair. In meiosis, Ser-140 phosphorylation (H2AX139ph) may occur at synaptonemal complexes during leptotene as an ATM-dependent response to the formation of programmed DSBs by SPO11. Ser-140 phosphorylation (H2AX139ph) may subsequently occurs at unsynapsed regions of both autosomes and the XY bivalent during zygotene, downstream of ATR and BRCA1 activation. Ser-140 phosphorylation (H2AX139ph) may also be required for transcriptional repression of unsynapsed chromatin and meiotic sex chromosome inactivation (MSCI), whereby the X and Y chromosomes condense in pachytene to form the heterochromatic XY-body. During immunoglobulin class switch recombination in lymphocytes, Ser-140 phosphorylation (H2AX139ph) may occur at sites of DNA-recombination subsequent to activation of the activation-induced cytidine deaminase AICDA. Phosphorylation at Tyr-143 (H2AXY142ph) by BAZ1B/WSTF determines the relative recruitment of either DNA repair or pro-apoptotic factors. Phosphorylation at Tyr-143 (H2AXY142ph) favors the recruitment of APBB1/FE65 and pro-apoptosis factors such as MAPK8/JNK1, triggering apoptosis. In contrast, dephosphorylation of Tyr-143 by EYA proteins (EYA1, EYA2, EYA3 or EYA4) favors the recruitment of MDC1-containing DNA repair complexes to the tail of phosphorylated Ser-140 (H2AX139ph).
Monoubiquitination of Lys-120 (H2AXK119ub) by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression. 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.
- Cellular localizationNucleus. Chromosome.
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Anti-Histone H2A.X antibody images
Lanes 1 - 2 : Anti-Histone H2A.X antibody (ab47503) at 1/500 dilution
Lanes 3 - 4 : Anti-Histone H2A.X (phospho S140) antibody (ab47371) at 1/500 dilution
Lane 1 : Extracts from Hela cells treated with UV
Lane 2 : Extracts from Hela cells treated with UV with blocking peptide
Lane 3 : Extracts from Hela cells
Lane 4 : Extracts from Hela cells treated with UV
Predicted band size : 15 kDa
Observed band size : 15 kDa
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ab47503 at a 1/50 dilution.
Left image : Untreated
Right image : Treated with Peptide
References for Anti-Histone H2A.X antibody (ab47503)
ab47503 has not yet been referenced specifically in any publications.