NF-kappaB p65 (Phospho-Ser311)Colorimetric Cell-Based ELISA Kit

Assay Genie's NF-kappaB p65 (Phospho-Ser311) Colorimetric Cell-Based ELISA Kit is a cell-based, lysate-free assay designed for the qualitative detection of phosphorylated NF-kappaB p65 at Ser311 in cultured adherent cells. The kit uses an indirect ELISA format in which target-specific primary antibodies capture the phospho-protein, and HRP-conjugated secondary antibodies generate a colorimetric signal at 450 nm. A monoclonal anti-GAPDH antibody is included as an internal positive control, and a Crystal Violet whole-cell stain can be used to normalise absorbance readings to cell density.

Reactivity has been characterised for human and mouse samples, with a dynamic range of > 5000 cells. The assay can be applied to measure relative phosphorylation levels in cultured cells and to screen the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on NF-kappaB p65 phosphorylation. The target protein corresponds to the RELA gene (Gene ID 5970; UniProt Q04206; chromosomal location 11q13.1), with a reported molecular weight of 59,910 Da, and is associated with inflammatory and immune signalling, including regulation of NF-kappaB transcription factor activity.