Chk1 (Phospho-Ser296)Colorimetric Cell-Based ELISA Kit

Assay Genie offers a cell-based ELISA kit for the qualitative detection of phosphorylated Chk1 at Ser296 in cultured cells. The kit uses an indirect ELISA format in which Chk1 (Phospho-Ser296) is captured by target-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, allowing the conjugated HRP enzyme to catalyze a colorimetric reaction upon substrate addition. This lysate-free, high-throughput assay is suitable for measuring relative Chk1 phosphorylation levels in cultured cells and for screening the effects of various treatments, inhibitors, or activators on Chk1 phosphorylation.

Detection is colorimetric, measured at 450 nm, with a stated dynamic range of > 5000 cells. Reactivity is reported for human samples, and the kit is intended for use with adherent cells. Normalization of the qualitative readout is supported by two methods: a monoclonal antibody specific for human GAPDH, included as an internal positive control for normalizing target absorbance values, and a Crystal Violet whole-cell staining procedure for determining cell density so that absorbance values can be adjusted for plating differences.

Chk1 is a serine/threonine-protein kinase required for checkpoint-mediated cell cycle arrest and activation of DNA repair in response to the presence of DNA damage or unreplicated DNA. Phosphorylated Chk1 at Ser296 plays a key role in cell cycle checkpoint control and DNA repair mechanisms, and dysregulation of this phosphorylation has been associated with diseases including cancer and genetic disorders, making the target relevant to therapeutic research and drug development. The Chk1 entry is identified in the source as Gene ID 1111 (UniProt ID O14757, OMIM 603078, Unigene Hs.24529) under the gene symbol CHK1 (also CHEK1), with a reported molecular weight of 50,415 Da.