KAPC A/B Colorimetric Cell-Based ELISA

From Assay Genie, this in-cell ELISA kit is designed for the direct detection of KAPC A/B protein expression in cultured cells without the need for lysate preparation. Operating on a 96-well microplate format with adherent cells, the cell-based, high-throughput, and sensitive assay provides qualitative measurement of KAPC A/B through an indirect ELISA format: KAPC A/B-specific primary antibodies are captured on the plate, and HRP-conjugated secondary antibodies then catalyse a colorimetric reaction at 450 nm upon substrate addition. The kit is reactive with human, mouse, and rat samples and has a dynamic range of > 5000 cells. It is suitable both for measuring relative KAPC A/B levels in cultured cells and for screening the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on KAPC A/B expression.

For data normalisation, a monoclonal antibody specific for human GAPDH is included as an internal positive control, and following the colorimetric read, a Crystal Violet whole-cell staining method can be used to determine cell density, allowing absorbance values to be normalised to cell amounts and correcting for plating differences. The target corresponds to PRKACA, the catalytic subunit of cAMP-dependent protein kinase alpha. Reagents are stored at 4°C with a stated stability of 6 months.