Smad2 (Phospho-Ser250)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie designed to measure Smad2 protein phosphorylation and expression profile in cultured adherent cells, specifically detecting Smad2 (Phospho-Ser250). The kit uses a colorimetric detection method at 450 nm in an indirect ELISA format, in which Smad2 (Phospho-Ser250)-specific primary antibodies capture the target and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody to catalyse a colour reaction upon substrate addition. This lysate-free workflow allows the relative amounts of phosphorylated Smad2 to be measured directly in fixed cells, making it suitable for screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on Smad2 phosphorylation.

Reactivity is reported for human, mouse, and rat samples, with a stated dynamic range of > 5000 Cells. The kit includes a monoclonal anti-GAPDH antibody to serve as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step is supplied to determine cell density so that plating differences can be adjusted for. The kit is supplied with the buffers, anti-phospho and anti-target antibodies, anti-GAPDH antibody, HRP-conjugated anti-rabbit and anti-mouse IgG antibodies, SDS solution, stop solution, ready-to-use substrate, crystal violet solution, and adhesive plate seals required to perform the assay.

Phosphorylation of Smad2 at serine 250 is described as a critical event in the TGF-β signalling pathway, regulating cellular processes including cell growth, differentiation, and apoptosis, with dysregulation of Smad2 phosphorylation implicated in cancer, fibrosis, and developmental disorders. The target corresponds to human SMAD2 (Gene ID: 4087, UniProt: Q15796, OMIM: 601366), making the kit a relevant tool for cell biology and signal transduction research into the TGF-β pathway and its role in disease pathogenesis.