IKK-beta (Phospho-Tyr188)Colorimetric Cell-Based ELISA Kit

A high-quality cell-based ELISA kit from Assay Genie designed for the detection of IKK-beta phosphorylation at Tyr188 in cell lysates. The assay uses a colorimetric detection method with absorbance measurement at 450 nm and has been validated for reactivity with Human, Mouse, and Rat samples. The kit has a dynamic range of > 5000 Cells and is described as a convenient, lysate-free, high-throughput, and sensitive assay.

Detection is achieved through an indirect ELISA format, in which IKK-beta (Phospho-Tyr188) is captured by specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. The conjugated HRP enzyme catalyses a colorimetric reaction upon substrate addition, enabling qualitative determination of phosphorylated IKK-beta in cultured cells. The protocol supports measuring the relative amounts of phosphorylated IKK-beta, as well as screening the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on IKK-beta phosphorylation.

Because the assay is qualitative, two normalization methods are provided: a monoclonal antibody specific for human GAPDH is included as an internal positive control to normalize target absorbance values, and a Crystal Violet whole-cell staining method can be used to determine cell density, allowing absorbance values to be adjusted to cell amounts.