The transcription factor NFκB (nuclear factor κB) is a key component for the inducible expression of a wide variety of cellular and viral genes. NFκB is composed of a heterodimer of p65 and p50 subunits in most cell types and is sequestered in the cytoplasm by its inhibitory proteins, the IκBs. During the phosphorylation and degradation of IκBs, NFκB p65 is phosphorylated on multiple resides, each triggered by different stimuli but essential for maintaining NFκB transcriptional activation. Because of its role in regulating inflammatory and immune responses, high-throughput study methods for monitoring NFκB phosphorylation are in high demand.

