This assay system utilizes HEK293t cells that express NF-κB (nuclear factor kappa-lightchain enhancer of activated B cells) and contain the luciferase reporter gene functionally linked to upstream NF-κB genetic response elements. Thus, quantifying changes in luciferase expression provides a sensitive surrogate measure of changes in the level of NF-κB activation. NF-κB is a signal transduction dependent transcription factor. This NF-κB reporter cell line is validated to provide a robust dose-dependent activation response when treated with TNFɑ, or the Protein Kinase C activator Phorbol 12-myristate 13-acetate (PMA). As such, the principle application of this assay system is in the screening of test samples to quantify any functional activities that they may exert to modulate, either induce or suppress, NF-κB activities. The assay kits are all-inclusive cell-based assay systems. In addition to NF-κB Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the users test samples, a positive control activator of NF-κB, Luciferase Detection Reagent, and a cell culture-ready assay plate.
▪ NF-κB Reporter Cells: 1 x 2.0 mL; -80°C▪ Cell Recovery Medium (CRM): 2 x 10.5 mL; -20°C▪ Compound Screening Medium (CSM): 1 x 45 mL; -20°C ▪ PMA*, 30 µM (in DMSO) (positive control for NF-κB activation via PKC pathways): 1 x 30 µL; -20°C▪ Detection Substrate: 1 x 6.0 mL; -80°C ▪ Detection Buffer: 1 x 6.0 mL; -20°C ▪ 96-well, collagen-coated assay plate (white, sterile, cell-culture ready): 1; -20°C