Chk1 (Phospho-Ser301)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed for the qualitative detection of Chk1 protein phosphorylated at Ser301 in cultured cells. The assay uses a colorimetric method at 450 nm in an indirect ELISA format, allowing measurement of relative levels of phosphorylated Chk1 and screening of treatments, inhibitors (such as siRNA or chemicals), or activators that affect Chk1 phosphorylation. The dynamic range is > 5000 Cells, and reactivity covers human, mouse, and rat samples. Sample type is adherent cells.

A monoclonal anti-GAPDH antibody is included to serve as an internal positive control for normalizing target absorbance values. Following the HRP-based colorimetric readout, a Crystal Violet whole-cell staining step may be used to determine cell density, so that absorbance values can be normalized to cell amounts and plating differences adjusted. In the workflow, Chk1 (Phospho-Ser301) is captured by target-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody and catalyze a colorimetric reaction upon substrate addition.

Chk1 is a serine/threonine kinase required for checkpoint-mediated cell cycle arrest and activation of DNA repair in response to the presence of DNA damage or unreplicated DNA. Phosphorylation at Ser301 is a marker relevant to Chk1 activation and function, making this kit applicable to research on DNA damage signaling, cell cycle regulation, and studies of potential therapeutic targets.