IKK-gamma Colorimetric Cell-Based ELISA Kit

This In-Cell ELISA Kit from Assay Genie is designed for the qualitative detection of IKK-gamma protein expression in cultured cells. The assay enables researchers to measure relative IKK-gamma levels in cell lysates and to screen the effects of various treatments, inhibitors (including siRNA or chemicals), or activators on IKK-gamma expression.

The kit uses a colorimetric detection method with absorbance read at 450 nm and is configured for a 96-well microplate format. It is reactive with human samples and is suitable for adherent cells, with a dynamic range of > 5000 cells. Storage is at 4 °C for 6 months. IKK-gamma, encoded by the IKBKG gene, is a regulatory subunit of the IKK core complex that phosphorylates inhibitors of NF-kappa-B, leading to dissociation of the inhibitor/NF-kappa-B complex and degradation of the inhibitor.

Detection follows an indirect ELISA format: IKK-gamma is captured by IKK-gamma-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. Following substrate addition, HRP catalyses a colour reaction. Because the assay is qualitative, two normalisation methods are required. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values. In addition, the Crystal Violet whole-cell staining method can be used to determine cell density, allowing absorbance values to be normalised to cell amounts and adjusting for plating differences.