Smad3 (Phospho-Ser425)Colorimetric Cell-Based ELISA Kit

A Cell-Based ELISA Kit from Assay Genie is designed for the accurate detection of phosphorylated Smad3 at the Serine 425 residue in cell lysates. The colorimetric assay is read at 450 nm and is reactive with Human, Mouse, and Rat samples, supporting a dynamic range of > 5000 Cells. It is intended for use with adherent cells and is classified as a phospho-specific Cell-Based ELISA Kit within the Immunology research area.

Working on a lysate-free, high-throughput format, the kit measures relative Smad3 phosphorylation and expression in cultured cells via an indirect ELISA approach. Target-specific primary antibodies capture Smad3 (Phospho-Ser425), and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalising target absorbance values.

Following the colorimetric read-out, the Crystal Violet whole-cell staining method may be applied to determine cell density, allowing results to be normalised to cell amounts and plating differences to be adjusted. The kit can be used to measure relative amounts of phosphorylated Smad3 in cultured cells and to screen for the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on Smad3 phosphorylation.