NF-kB p65 (Phospho-Ser529)Fluorometric Cell-Based ELISA Kit

This Fluorometric Cell-Based ELISA Kit from Assay Genie is designed to monitor NF-kB p65 (Phospho-Ser529) protein phosphorylation and expression profile in cultured cells. The lysate-free, high-throughput assay provides a sensitive approach for measuring the relative amounts of phosphorylated NF-kB p65 in cells, and can be used to screen the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on phosphorylation.

Detection is achieved through an indirect ELISA format that qualitatively determines NF-kB p65 (Phospho-Ser529) levels. Primary antibodies specific for the target capture the phosphorylated protein, while Dye 1- and Dye 2-conjugated secondary antibodies bind the primary antibody's Fc region, enabling fluorometric readout. Two normalization methods are included: a monoclonal antibody specific for human GAPDH serves as an internal positive control, and an antibody against the non-phosphorylated counterpart of NF-kB p65 (Phospho-Ser529) is provided so that relative fluorescence units obtained for non-phosphorylated NF-kB p65 can be used to normalize the value for the phosphorylated target.

The kit has a dynamic range of > 5000 cells and is reactive with human, mouse, and rat samples. Storage is at 4°C with 6 months stability. The associated NCBI gene symbol is RELA (Gene ID: 5970; UniProt ID: Q04206), and the intended sample type is cells.