Smad1/5/9 Colorimetric Cell-Based ELISA

This in-cell ELISA kit from Assay Genie detects and quantifies Smad1, Smad5 and Smad9 protein expression profiles directly in cultured adherent cells, using a colorimetric readout at 450 nm. The lysate-free workflow employs an indirect ELISA format, where target-specific primary antibodies are captured on the cells and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody to catalyze a colorimetric reaction upon substrate addition.

The kit is reactive with human, mouse and rat samples and is suited to measuring relative amounts of Smad1/5/9 in cultured cells, as well as screening the effects of treatments, inhibitors (such as siRNA or chemicals) or activators on Smad1/5/9 levels. Because the assay is qualitative, two normalization methods are provided: a monoclonal anti-GAPDH antibody serves as an internal positive control for normalizing target absorbance values, and Crystal Violet whole-cell staining is used to determine cell density so absorbance values can be adjusted to cell amount.

Key specifications include a dynamic range of > 5000 cells per well, a 96-Well Microplate format and storage at 4 °C with 6 months stability.