Smad3 (Phospho-Ser208)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit enables measurement of phosphorylated Smad3 (Ser208) levels in cultured cells without the need for lysate preparation. The assay uses a colorimetric detection method with absorbance read at 450 nm, providing a high-throughput, sensitive means of profiling Smad3 phosphorylation and expression in cells.

The kit is reactive with human, mouse and rat samples, with a dynamic range of greater than 5000 cells, and is intended for adherent cell samples. Detection is achieved through an indirect ELISA format, in which Smad3 (Phospho-Ser208) is captured by specific primary antibodies and detected using HRP-conjugated secondary antibodies that catalyze a colorimetric reaction upon substrate addition.

To account for the qualitative nature of cell-based ELISA, the kit includes a monoclonal anti-GAPDH antibody as an internal positive control for normalizing target absorbance, along with a Crystal Violet whole-cell staining method for determining cell density. Researchers can use the kit to measure relative Smad3 phosphorylation in cultured cells and to screen the effects of various treatments, inhibitors (such as siRNA or chemicals) or activators on Smad3 phosphorylation. SMAD3 is a key signaling molecule in the TGF-β pathway, with phosphorylation at Ser208 modulating its activity, and the kit is suited to research on TGF-β signaling, cell proliferation and differentiation, as well as diseases linked to dysregulated TGF-β signaling such as cancer, fibrosis and developmental disorders.