NF-kappaB p105/p50 Transcription Factor Activity Assay

The Assay Genie NF-kappaB p105/p50 Transcription Factor Activity Assay quantifies DNA-binding activity of the NF-kB p50 subunit (generated by proteolytic processing of the p105 precursor, NF-kB1) in nuclear and cell lysates. The microplate is pre-coated with an immobilized double-stranded oligonucleotide containing the NF-kB consensus binding motif. Activated p50 in nuclear extracts binds the captured probe and is detected with a subunit-specific primary antibody and HRP-conjugated secondary reagent, with absorbance read on a standard plate reader.

The assay uses a sample volume of 5 to 20 uL per well and includes competitor oligonucleotide along with wild-type and mutant consensus controls to confirm binding specificity. The kit accommodates up to 96 individual sample determinations and ships with all required buffers and a TMB-based development substrate. Recommended for studies of inflammatory signaling, cytokine-driven gene expression, and inhibitor screening workflows in human, mouse, and rat models. Store components at -20 degrees C; avoid repeated freeze-thaw cycles of the antibody and probe reagents.