IkappaB-beta (Phospho-Ser23)Colorimetric Cell-Based ELISA Kit

Assay Genie provides a Cell-Based ELISA Kit for the qualitative detection of IkappaB-beta phosphorylated at Ser23 in cultured cells. The assay is lysate-free and uses an indirect ELISA format, in which phospho-specific primary antibodies capture the target while HRP-conjugated secondary antibodies catalyze a colorimetric reaction measured at 450 nm. The kit is reactive with human, mouse, and rat samples and is intended for adherent cells, supporting work on NF-kB pathway activation, inflammation, immune response, and cell survival. A dynamic range of > 5000 cells enables high-throughput screening of the effects that various treatments, inhibitors (e.g., siRNA or chemicals), or activators have on IkappaB-beta phosphorylation status.

To control for inter-well variation, the kit includes a monoclonal anti-GAPDH antibody for internal positive-control normalization, along with a Crystal Violet whole-cell staining method to determine cell density after the colorimetric read. The associated target, NFKBIB (Gene ID: 4793; UniProt ID: Q15653), encodes an NF-kappa-B inhibitor family member that retains NF-kB in the cytoplasm; phosphorylation of serine residues on these proteins marks them for destruction, freeing NF-kB to translocate to the nucleus and function as a transcription factor. The kit is therefore suited to studies of signaling pathway activation and the identification of potential therapeutic targets in conditions such as cancer, autoimmune disorders, and inflammatory diseases.