Kit (Phospho-Tyr721)Colorimetric Cell-Based ELISA Kit

An In-Cell ELISA kit from Assay Genie enables direct detection of Kit phosphorylation at tyrosine 721 in cultured adherent cells, without the need for lysate preparation. The assay uses an indirect ELISA format in which Kit (Phospho-Tyr721)-specific primary antibodies capture the target, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody to generate a colorimetric signal at 450 nm upon substrate addition. With a dynamic range of greater than 5000 cells, the kit is validated for use with Human, Mouse, and Rat samples and is intended for measuring relative levels of phosphorylated Kit in cultured cells as well as for screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on Kit phosphorylation.

Two normalisation approaches are provided to account for the qualitative nature of the cell-based ELISA: a monoclonal antibody specific for human GAPDH serves as an internal positive control for normalising target absorbance values, while a Crystal Violet whole-cell staining method may be used to determine cell density, enabling absorbance values to be normalised to cell amounts and correct for plating differences.