Smad2/3 (Phospho-Thr8)Colorimetric Cell-Based ELISA Kit

The Assay Genie cell-based ELISA kit enables quantitative detection of phosphorylated SMAD2/3 at Thr8 in cell lysates. Using an indirect ELISA format, Smad2/3 (Phospho-Thr8) is captured by target-specific primary antibodies while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction upon substrate addition. The assay is read at 450 nm and has a stated dynamic range of > 5000 Cells.

Reactivity is established for Human, Mouse and Rat samples, and the procedure is designed for adherent cells. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and the Crystal Violet whole-cell staining method may be used to determine cell density so that absorbance values can be normalised to cell amounts.

SMAD2 and SMAD3 are key regulators of TGF-β signalling, controlling processes such as growth, differentiation and apoptosis, and phosphorylation at Thr8 is essential for their activation and nuclear translocation to regulate gene expression. The phospho-specific kit is classified for use in the Immunology research area and is suited to studying SMAD2/3 signalling in the context of diseases such as cancer, fibrosis and developmental disorders.