FIR Colorimetric Cell-Based ELISA

A Cell-Based ELISA from Assay Genie that provides a lysate-free, high-throughput method for detecting FIR protein expression in cultured cells. The assay uses an indirect ELISA format in which FIR is captured by target-specific primary antibodies; HRP-conjugated secondary antibodies then bind the Fc region of those primaries, and the conjugated HRP enzyme catalyses a colorimetric reaction upon substrate addition.

The kit is suited to measuring relative amounts of FIR in cultured cells and to screening the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on FIR. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step is used to determine cell density, allowing absorbance readings to be normalised to cell amounts.

Reactivity is reported for human and mouse, with a dynamic range of > 5000 cells and colorimetric detection at 450 nm. The assay is supplied on a 96-well microplate format, is intended for use with adherent cells, and is stored at 4 °C with a stated stability of 6 months. FIR is also known as FARP2 (FERM, ARHGEF and pleckstrin domain-containing protein 2), a Rho-guanine nucleotide exchange factor that activates RAC1, with a reported molecular weight of 119 kDa (UniProt O94887).