| H2AX comprises approximately 2-25% of the histone H2A pool in mammalian cells. The extended C-terminal tail of H2AX contains a serine (Ser139) embedded in an conserved SQE motif, which is rapidly phosphorylated at sites of double strand breaks and stalled replication forks. Phosphorylated H2AX may serve to recruit other repair proteins to the site of DNA damage. |
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| H2AX has been implicated in both homologous recombination and nonhomologous end joining DNA repair pathways. |
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| Catalog # | H5110-08C |
| Applications | Suitable for use in Western Blot, Immunofluorescence (2), Chromatin Immunoprecipitation (2) and Immunocytochemistry. Other applications not tested. |
| Recommended Dilution | Western Blot Analysis: 0.5-1ug/ml detects phosphorylated histone H2A.X (Ser139) in acid extracted histone from Jurkat cell lysates treated with 0.5uM staurosporine. Untreated, and staurosporine-treated Jurkat cell lysates were resolved by electrophoresis, transferred to nitrocellulose and probed with H5110-08C (0.5ug/ml). Proteins were visualized using a IgG (HRP) GtxMo and a chemiluminescence detection system. |
| Immunocytochemistry: 2ug/ml detects phosphorylated histone H2A.X (Ser139) in HeLa cells treated with 0.5uM staurosporine for 4-6 hours. |
| Optimal dilutions to be determined by the researcher. |
| Recommended secondary antibodies | I1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo |
| I1904-06H IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo |
| Positive Control | UV-treated 293 cell exxtracts, UV-treated HeLa cell extracts, breast cancer tissue. |
| Storage and Stability | May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| CAS Number | n/a |
| Clone Type | Monoclonal |
| Isotype | IgG1 |
| Clone No | 2T14 |
| Host | Mouse |
| Source | Human |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in PBS, pH 7.6, 0.07% sodium azide and 30% glycerol. |
| Purity | Purified by Protein G affinity chromatography. |
| Immunogen | Synthetic peptide (C-KATQA[pS]QEY) corresponding to aa134-142 of human histone H2A.X. (KLH). The immunizing sequence has 8 identical amino acids in yeast and mouse. MW: 17kD |
| Specificity | Recognizes human Histone H2A.X phosphorylated at Ser139, Mr 15kD. A broad species crossreactivity is expected based on conservation of sequence homology. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
