IkappaB-epsilon (Phospho-Ser22)Colorimetric Cell-Based ELISA Kit

Designed for the qualitative detection of IkappaB-epsilon (Phospho-Ser22) phosphorylation and expression in cultured cells, this Cell-Based ELISA Kit from Assay Genie is a lysate-free, high-throughput, and sensitive assay. It uses an indirect ELISA format, in which target-specific primary antibodies capture the analyte and HRP-conjugated secondary antibodies catalyze a colorimetric reaction upon substrate addition. The kit is reactive with Human, Mouse, and Rat samples and uses a colorimetric detection method measured at 450 nm, with a dynamic range of > 5000 Cells.

The workflow supports relative measurement of phosphorylated IkappaB-epsilon in adherent cell cultures and is suitable for screening the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on IkappaB-epsilon phosphorylation. Two normalization strategies are provided: a monoclonal anti-GAPDH antibody serves as an internal positive control for normalizing target absorbance, and a Crystal Violet whole-cell staining step can be used to determine cell density and adjust for plating differences. The target protein, IkappaB-epsilon (gene symbol NFKBIE, UniProt ID: O00221), is a member of the NF-kappa-B inhibitor family that sequesters NF-kappa-B complexes in the cytoplasm.