Histone H2A.X, phosphorylated (Ser139/Tyr142) (H2a/x, H2AFX, H2AX)

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 on 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 on 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). \n\nApplications: \nSuitable for use in Immunofluorescence, Flow Cytometry, Western Blot and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution:\nImmunofluorescence (IC): 1:100 \nFlow Cytometry: 1:200\nWestern Blot: 1:1000 \nImmunoprecipitation: 1:50\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts

SPECIFICATIONS

Catalog Number

H5110-03W

Size

100ul

Applications

FC, IF, IP, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat, NHP

Form

Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

P Type

Pab

Purity

Purified by Protein A and peptide affinity chromatography.

Isotype

IgG

References

1.Yuan, J. et al. (2010) FEBS Lett 584, 3717-24. 2.Rogakou, E.P. et al. (1998) J Biol Chem 273, 5858-68. 3.Burma, S. et al. (2001) J Biol Chem 276, 42462-7. 4.Rogakou, E.P. et al. (1999) J Cell Biol 146, 905-16. 5.Mukherjee, B. et al. (2006) DNA Repair (Amst) 5, 575-90. 6.Solier, S. et al. (2009) Mol Cell Biol 29, 68-82. 7.Lu, C. et al. (2006) Mol Cell 23, 121-32. 8.Lu, C. et al. (2008) FEBS Lett 582, 2703-8. 9.Cook, P.J. et al. (2009) Nature 458, 591-6. 10.Xiao, A. et al. (2009) Nature 457, 57-62.

Additional Info

Recognizes endogenous levels of human histone H2A.X, phosphorylated at Ser139 or Tyr142, or dually phosphorylated on both residues. Does not react with Other applications have not been tested. histone proteins. Species Crossreactivity: monkey, mouse and rat.