NF-kappaB p105/p50 (Phospho-Ser893)Colorimetric Cell-Based ELISA Kit

A cell-based, lysate-free colorimetric ELISA from Assay Genie enables qualitative detection of NF-kappaB p105/p50 phosphorylated at Ser893 in cultured cells. The kit uses an indirect ELISA format: a phospho-specific primary antibody captures the target, and an HRP-conjugated secondary antibody binds the Fc region of the primary, producing a colorimetric signal upon substrate addition that is read at 450 nm. The assay has a dynamic range of > 5000 Cells and is validated for use with human samples and adherent cells.

To account for the qualitative nature of the cell-based ELISA, two normalization strategies are included. A monoclonal anti-human GAPDH antibody serves as an internal positive control for normalizing target absorbance, and Crystal Violet whole-cell staining is provided for determining cell density so that absorbance values can be normalized to cell number, correcting for plating variation. The kit is suited to measuring relative phosphorylation levels in response to treatments, inhibitors (including siRNA or chemicals), and activators.

NF-kappaB is a key transcription factor that regulates immune response, inflammation, and cell survival, and phosphorylation at Ser893 is reported to modulate its activity. The target is the NFKB1 gene product (UniProt P19838), a DNA-binding transcription factor. Applications include research into inflammatory diseases, cancer biology, and signal transduction pathways.