NF-kappaB p105/p50 (Phospho-Ser927)Colorimetric Cell-Based ELISA Kit

A colorimetric cell-based ELISA kit from Assay Genie is designed for the precise detection of phosphorylated NF-kappaB p105/p50 (Phospho-Ser927) levels in cell lysates. The assay is reactive with human and mouse samples and has a stated dynamic range of > 5000 Cells. Phosphorylation of NF-kappaB p105/p50 at Ser927 plays a crucial role in the activation of NF-kappaB signaling, making it a vital marker for studies of immune and inflammatory responses, cell survival and proliferation, as well as research into cancer, inflammation, and autoimmune disorders.

The kit is intended for use with adherent cells and employs an indirect ELISA format in which the target is captured by phospho-specific primary antibodies and detected using HRP-conjugated secondary antibodies that catalyze a colorimetric reaction at 450 nm upon substrate addition. A monoclonal anti-human GAPDH antibody is included as an internal positive control for normalization, and the Crystal Violet whole-cell staining method can be used to determine cell density, after which absorbance values are normalized to cell amounts to adjust for plating differences. The assay can be used to measure relative amounts of phosphorylated NF-kB p105/p50 in cultured cells and to screen for the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on NF-kB p105/p50 phosphorylation.

According to the associated UniProt record, the NFKB1 gene encodes an 85,520 Da DNA-binding transcription factor (NFkB-p105) that undergoes cotranslational processing by the 26S proteasome to produce a 50 kD DNA-binding subunit. NFkB is activated by diverse intra- and extracellular stimuli such as cytokines, oxidant-free radicals, ultraviolet irradiation, and bacterial or viral products, and translocates into the nucleus to stimulate expression of genes involved in a wide variety of biological functions, including inflammatory and innate immune responses and apoptosis.