CDK1/CDC2 (Phospho-Thr14)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this colorimetric cell-based ELISA kit is designed for researchers investigating phosphorylation of CDK1/CDC2 at threonine 14. It provides a lysate-free, high-throughput and sensitive approach for detecting CDK1/CDC2 protein phosphorylation and expression profiles directly in cultured cells, and can be used to measure the relative amounts of phosphorylated CDK1/CDC2 in cultured cells as well as to screen for the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on CDK1/CDC2 phosphorylation.

The assay follows an indirect ELISA format in which the target is captured by CDK1/CDC2 (Phospho-Thr14)-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, enabling a colorimetric readout upon substrate addition. The detection wavelength is 450 nm and the dynamic range is > 5000 cells. Reactivity is established for human, mouse and rat samples, and the kit is suited to adherent cells.

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 step may be used to determine cell density, allowing absorbance values to be normalised to cell amounts and plating differences to be adjusted. Qualitative determination of CDK1/CDC2 (Phospho-Thr14) concentration is achieved, supporting studies of cell cycle regulation and potential therapeutic targets.