Smad3 (Phospho-Ser213)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed to detect and quantify Smad3 phosphorylated at Ser213 in cultured cells. It is a lysate-free, high-throughput indirect ELISA in which the phospho-Smad3 target is captured by specific primary antibodies and revealed with HRP-conjugated secondary antibodies that catalyze a colorimetric reaction read at 450 nm. Reactivity is reported for human, mouse and rat samples, and the kit is intended for use with adherent cells, with a dynamic range of > 5000 cells.

Because the assay is qualitative, two normalisation strategies are built in: a monoclonal anti-GAPDH antibody serves as an internal positive control for target absorbance values, and a Crystal Violet whole-cell staining step can be used to determine cell density and adjust for plating differences. The kit is used to measure the relative levels of phosphorylated Smad3 in cultured cells and to screen the effects of treatments, inhibitors (such as siRNA or chemicals) and activators on Smad3 phosphorylation. SMAD3 is a key signalling protein in the TGF-beta pathway, and phosphorylation at Ser213 is associated with its activation and downstream effects on gene expression, making the kit relevant to research on cell growth, differentiation, apoptosis, cancer, fibrosis and inflammation.