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H5110-03W Rabbit Anti-Histone H2A.X, phosphorylated (Ser139/Tyr142) (H2a/x, H2AFX, H2AX)

Specifications
References
Clone Type
Polyclonal
Host
Rabbit
Source
Human
Swiss Prot
P16104
Isotype
IgG
Grade
Affinity Purified
Applications
FC IF IP WB
Crossreactivity
Hu Mk Mo Rt
Gene ID
3014
Shipping Temp
Blue Ice
Storage Temp
-20°C

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).

Applications
Suitable for use in Immunofluorescence, Flow Cytometry, Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilution
Immunofluorescence (IC): 1:100 Flow Cytometry: 1:200 Western Blot: 1:1000 Immunoprecipitation: 1:50 Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Synthetic phosphopeptide corresponding to residues surrounding the phosphorylated Ser139 and Tyr142 of human H2A.X.
Form
Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.
Purity
Purified by Protein A and peptide affinity chromatography.
Specificity
Recognizes endogenous levels of human histone H2A.X, phosphorylated at Ser139 or Tyr142, or dually phosphorylated on both residues. Does not react with histone proteins. Other applications have not been tested. Species Crossreactivity: monkey, mouse and rat.
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.
USBio References
No references available
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