Research detection kit HTRF

cell

The Total H2AX kit is designed to monitor the expression level of cellular human H2AX, and can be used as a normalization assay for the Phospho-H2AX (Ser139) Detection kit. OverView The Total-H2AX kit offers quantitative detection of human histone variant H2AX. The specific phosphorylation of H2AX at Ser139 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, which makes this assay particularly appropriate for studying genome stability, cell cycle, DNA repair, and more broadly as a biomarker for cancer and for aging process investigations. Other Specifications Unit Size 500 Assay Points How it works Total H2AX assay principle The Total-H2AX assay quantifies the expression level of H2AX in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-H2AX assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of H2AX in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein’s expression under a no-wash assay format.