H2A histone family member X (H2AX) is a type of histone protein encoded by the H2AFX gene. H2AX phosphorylation on serine 139 may occur when double-stranded DNA breaks, either due to ionizing radiation or to endogenous physiological processes. This modification, called gamma-H2AX (phospho-Ser139), triggers cell cycle arrest and DNA damage response repair. Analyzing H2AX allows for a better understanding of genome stability, cell cycle, DNA repair, and more broadly, H2AX is a biomarker for cancer and for aging process investigations.

The AlphaLISA SureFire Ultra Multiplex Human Phospho-H2A.X (Ser139)/Total H2A.X Detection Kit is used to measure phosphorylated H2A.X (Ser139) and total levels of H2A.X in cellular lysates in a multiplexing Alpha no-wash assay. No specially engineered cell lines are required. The 615 nm (Eu) signal corresponds to the phosphorylated H2A.X, and the 545 nm (Tb) signal corresponds to the total H2A.X analysis. This kit has been formulated to provide superior signal:background assay windows and to perform without interference in the presence of extraneous antibodies, making it amenable to the study of therapeutic and blocking antibodies.

• No-wash steps, no separation steps
• Multiplexing assay
• Sensitive detection
• No interference from exogenous antibodies
• Results in less than 3 hours

In this assay, the Alpha 615 Acceptor bead is coated with the CaptSure^™ antibody, which binds the CaptSure-tagged anti-phospho target antibody. The Alpha 545 Acceptor bead is coated with the CaptSure2^™ agent, which binds the CaptSure2 tagged anti-total target protein antibody. The Alpha Donor bead binds the biotinylated anti-total target protein antibody.