Histone H3 (Phospho-Ser10)Colorimetric Cell-Based ELISA Kit

A Cell-Based ELISA Kit from Assay Genie designed for the precise measurement of Histone H3 phosphorylation at serine 10 in cell samples. The kit uses a colorimetric detection method at 450 nm and is validated for reactivity with Human, Mouse, and Rat samples, with a dynamic range of > 5000 Cells.

Optimized as a convenient, lysate-free, high-throughput and sensitive assay, the platform employs an indirect ELISA format. Histone H3 (Phospho-Ser10) is captured by target-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. Upon substrate addition, the conjugated HRP enzyme catalyses a colorimetric reaction. The kit is intended for use with adherent cells and can be used both for measuring the relative amounts of phosphorylated Histone H3 in cultured cells and for screening the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on Histone H3 phosphorylation.

For data analysis, a monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing target absorbance values. Following the colorimetric measurement of HRP activity via substrate addition, the Crystal Violet whole-cell staining method may be used to determine cell density, allowing the absorbance values to be normalized to cell amounts and plating differences to be adjusted.