Smad3 Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie provides a convenient, lysate-free method for measuring Smad3 protein expression directly in cultured adherent cells. The kit uses an indirect ELISA format, in which Smad3 is captured by Smad3-specific primary antibodies and detected using HRP-conjugated secondary antibodies that catalyze a colorimetric reaction upon substrate addition. Researchers can use the kit to assess relative Smad3 levels in cell samples and to screen the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on Smad3.

The assay uses a high-throughput microplate format with colorimetric detection at 450 nm and has a stated dynamic range of more than 5000 cells per well. It reacts with human, mouse, and rat samples. Two normalization strategies are included: a monoclonal anti-GAPDH antibody serves as an internal positive control for standardizing target absorbance values, and a Crystal Violet whole-cell stain can be used to determine cell density so that absorbance readings can be normalized to cell number. The kit is stored at 4 °C and is reported to be stable for six months under those conditions.

Smad3 (also known as Mothers against decapentaplegic homolog 3, gene symbol SMAD3, NCBI Gene ID 4088, UniProt P84022) is a receptor-regulated SMAD that acts as an intracellular signal transducer and transcriptional modulator activated by TGF-β and activin type 1 receptor kinases. Aberrant SMAD3 activity has been linked to cancer, fibrosis, and autoimmune disorders, and the kit is categorized for use in immunology research. The Smad3 protein has a reported molecular weight of 25,722 Da.