Rabbit anti-Human H2A.x-S139 Antibody | anti-H2AFX antibody

Phospho-Histone H2A.x-S139 Polyclonal Antibody

Cat. Num. AAA126189

• Gene Names: H2AFX; H2AX; H2A.X; H2A/X
• Reactivity: Human
• Applications: Western Blot, Immunohistochemistry, Immunofluorescence
• Purity: Affinity Purification
• Synonyms: H2A.x-S139; Phospho-Histone H2A.x-S139 Polyclonal Antibody; Phospho-H2AFX-S139: H2AFX; Histone H2A.x; anti-H2AFX antibody

• Host: Rabbit
• Reactivity: Human
• Isotype: IgG
• Purity/Purification: Affinity Purification
• Concentration: 1mg/ml (varies by lot)
• Sequence Length: 143

• Applicable Applications for anti-H2AFX antibody: Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF)
• Application Notes: WB: 1:500 - 1:2000; IHC: 1:50 - 1:200; IF: 1:50 - 1:200
• Species: Human
• Route: Synthetic Peptide
• Immunogen: A phospho specific peptide corresponding to residues surrounding S139 of human Histone H2A.x
• Calculated Molecular Weight: 14kDa
• Preparation and Storage: Store at -20 degree C (regular) or -80 degree C (long term). Avoid freeze / thaw cycles.; Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.

Western Blot (WB)

(Western blot analysis of extracts of Jurkat cells, using Phospho-H2AFX-S139 antibody. Jurkat cells were treated by etoposide.Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (MBS128200) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% BSA.)

Immunofluorescence (IF)

(Immunofluorescence analysis of U2OS cells using Phospho-H2AFX-S139 antibody. Blue: DAPI for nuclear staining.)

Immunofluorescence (IF)

(Immunofluorescence analysis of U2OS cells using Phospho-H2AFX-S139 antibody. Blue: DAPI for nuclear staining.)

Related Product Information for anti-H2AFX antibody

Histone H2A.X is a variant histone that represents approximately 10% of the total H2A histone proteins in normal human fibroblasts (1). H2A.X is required for checkpoint-mediated cell cycle arrest and DNA repair following double-stranded DNA breaks (1). DNA damage, caused by ionizing radiation, UV-light, or radiomimetic agents, results in rapid phosphorylation of H2A.X at Ser139 by PI3K-like kinases, including ATM, ATR, and DNA-PK (2,3). Within minutes following DNA damage, H2A.X is phosphorylated at Ser139 at sites of DNA damage (4). This very early event in the DNA-damage response is required for recruitment of a multitude of DNA-damage response proteins, including MDC1, NBS1, RAD50, MRE11, 53BP1, and BRCA1 (1). In addition to its role in DNA-damage repair, H2A.X is required for DNA fragmentation during apoptosis and is phosphorylated by various kinases in response to apoptotic signals. H2A.X is phosphorylated at Ser139 by DNA-PK in response to cell death receptor activation, c-Jun N-terminal Kinase (JNK1) in response to UV-A irradiation, and p38 MAPK in response to serum starvation (5-8). H2A.X is constitutively phosphorylated on Tyr142 in undamaged cells by WSTF (Williams-Beuren syndrome transcription factor) (9,10). Upon DNA damage, and concurrent with phosphorylation of Ser139, Tyr142 is dephosphorylated at sites of DNA damage by recruited EYA1 and EYA3 phosphatases (9). While phosphorylation at Ser139 facilitates the recruitment of DNA repair proteins and apoptotic proteins to sites of DNA damage, phosphorylation at Tyr142 appears to determine which set of proteins are recruited. Phosphorylation of H2A.X at Tyr142 inhibits the recruitment of DNA repair proteins and promotes binding of pro-apoptotic factors such as JNK1 (9). Mouse embryonic fibroblasts expressing only mutant H2A.X Y142F, which favors recruitment of DNA repair proteins over apoptotic proteins, show a reduced apoptotic response to ionizing radiation (9). Thus, it appears that the balance of H2A.X Tyr142 phosphorylation and dephosphorylation provides a switch mechanism to determine cell fate after DNA damage.

Product Categories/Family for anti-H2AFX antibody

Phosphorylation

NCBI and Uniprot Product Information

• NCBI GI #: 121992
• NCBI GeneID: 3014
• UniProt Accession #: P16104; Q4ZGJ7; Q6IAS5
• Molecular Weight: 143
• NCBI Official Full Name: Histone H2AX
• NCBI Official Synonym Full Names: H2A histone family, member X
• NCBI Official Symbol: H2AFX
• NCBI Official Synonym Symbols: H2AX; H2A.X; H2A/X
• NCBI Protein Information: histone H2AX; H2AX histone; histone H2A.x
• UniProt Protein Name: Histone H2AX
• UniProt Gene Name: H2AFX
• UniProt Synonym Gene Names: H2AX; H2a/x
• UniProt Entry Name: H2AX_HUMAN

NCBI Description

Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif. [provided by RefSeq, Jul 2008]

Uniprot Description

H2AX: a histone that replaces conventional H2A in a subset of nucleosomes. 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. Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called 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. Phosphorylated on S139 by ATM and DNA-PK 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. Dephosphorylation of S139 by PP2A is required for DNA DSB repair. Apparently phosphorylated on Y143 by WSTF, determining the relative recruitment of either DNA repair or pro-apoptotic factors. H2AXpY142 favors the recruitment of pro-apoptotic factors APBB1 and JNK1. In contrast, dephosphorylation of pY143 by EYA phosphatases favors the recruitment of MDC1-containing DNA repair complexes to the tail of phosphorylated pS139. Monoubiquitination of K119 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'K63' linkages by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage.

Protein type: Helicase; DNA repair, damage; DNA-binding

Chromosomal Location of Human Ortholog: 11q23.3

Cellular Component: nucleoplasm; XY body; male germ cell nucleus; chromosome, telomeric region; condensed nuclear chromosome; nuclear chromatin; replication fork; nucleosome; nucleus

Molecular Function: protein binding; enzyme binding; DNA binding; histone binding; protein heterodimerization activity; damaged DNA binding

Biological Process: nucleosome assembly; positive regulation of DNA repair; meiotic cell cycle; double-strand break repair; DNA damage checkpoint; spermatogenesis; response to ionizing radiation; DNA repair; response to DNA damage stimulus; double-strand break repair via homologous recombination