RAD17 (Phospho-Ser645)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Colorimetric Cell-Based ELISA Kit is designed for the detection of phosphorylated RAD17 at Ser645 in cultured cells. It uses an indirect ELISA format in which target-specific primary antibodies capture the phosphorylated protein, and HRP-conjugated secondary antibodies catalyse a colorimetric reaction upon substrate addition. Absorbance is read at 450 nm, and the procedure is lysate-free and high throughput, suitable for adherent cells as well as other cell types.

The kit supports studies of the DNA damage response and cell cycle checkpoint signalling. RAD17 is described as a cell cycle checkpoint protein required for cell cycle arrest and DNA damage repair, which shares similarity with DNA replication factor C, binds to chromatin prior to DNA damage, and is phosphorylated by ATR after damage. Phosphorylation at Ser645 is associated with activation of RAD17 and with the DNA-damage-induced G2 arrest response, making the phospho-Ser645 site a useful biomarker for DNA repair research and potential therapeutic target studies.

Reactivity covers Human and Mouse samples. The dynamic range is stated as > 5000 Cells. Because the assay is qualitative, two normalisation methods are provided: a monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining method can be used to determine cell density so that absorbance values can be normalised to cell amounts. The kit can be used for measuring relative levels of phosphorylated RAD17 in cultured cells and for screening the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on RAD17 phosphorylation, in both basic research and drug discovery applications.