NF-kappaB p65 (Phospho-Ser276)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit enables researchers to study phosphorylation of NF-kappaB p65 at Ser276, a regulatory event central to inflammatory and immune signalling. The assay supports investigations into the role of NF-kappaB in cancer, inflammatory diseases, and autoimmune disorders by measuring relative levels of phosphorylated NF-kappaB p65 in cultured cells.

Detection uses a lysate-free, indirect ELISA format: a phospho-specific primary antibody captures the target, and an HRP-conjugated secondary antibody catalyses a colorimetric reaction read at 450 nm. The kit is validated for Human, Mouse, and Rat samples using adherent cells, with a dynamic range of more than 5000 cells. An anti-GAPDH monoclonal antibody is supplied as an internal positive control for normalising target absorbance, and a Crystal Violet whole-cell staining step allows further normalisation to cell density to correct for plating differences.

Beyond direct measurement, the kit can screen the effects of various treatments, inhibitors (such as siRNA or chemicals), and activators on NF-kappaB p65 phosphorylation. The target is encoded by the RELA gene (Gene ID: 5970; UniProt ID: Q04206; OMIM: 164014), and the assay is run in a microplate-based format.