NF-kappaB p65 (Phospho-Ser536)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA from Assay Genie is designed to measure relative levels of NF-kappaB p65 phosphorylated at Ser536 in cultured adherent cells. The colorimetric assay is performed at 450 nm and is reactive with human, mouse and rat samples, with a dynamic range of > 5000 Cells. The kit is provided in a 96-well microplate format and uses a lysate-free, indirect ELISA workflow in which target-specific primary antibodies are paired with HRP-conjugated secondary antibodies, producing a colorimetric signal upon substrate addition.

Designed for high-throughput screening, the assay supports measurement of phosphorylation changes in response to treatments, inhibitors (such as siRNA or chemicals) or activators. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalising target absorbance, and a Crystal Violet whole-cell staining step allows results to be normalised to cell number, compensating for plating differences.

NF-kappaB p65 (gene symbol RELA) is a key transcription factor involved in regulating inflammatory and immune responses, with phosphorylation at Ser536 linked to its activation and nuclear translocation. Aberrant NF-kappaB signalling has been implicated in cancer, inflammatory disorders and autoimmune conditions, supporting the relevance of this phospho-specific assay to studies of disease mechanisms and potential therapeutic targets.