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Applications: Monitor NF-κB-signaling pathway activity Screen inhibitors of NF-κB
Description: The NF-κB Reporter Cellular Assay Pack provides all the key reagents required to monitor the activity of the nuclear factor Kappa B (NF-κB) signal transduction pathways. The pack contains the NF-κB Reporter (Luc)- CHO-K1 Recombinant Cell Line, a luciferase reporter cell line that contains a firefly luciferase gene under the control of four copies of the NF-κB response element located upstream of the minimal TATA promoter. After activation by pro-inflammatory cytokines or stimulants of lymphokine receptors, endogenous NF-κB transcription factors bind to the DNA response elements, inducing transcription of the luciferase reporter gene. This cell line is validated for the response to TNFalpha and to treatment with NF-κB inhibitor, evodiamine. Additionally, the pack includes cell culture medium (Thaw Medium 3) that has been optimized for use with CHO-K1 cells. Thaw Medium 3 includes 10% fetal bovine serum and 1% Pen/Strep. Finally, the pack provides the ONE-Step™ Luciferase Detection System. This reagent provides highly sensitive, stable detection of firefly (Photinus pyralis) luciferase activity. The ONE-Step luciferase reagent can be used directly in cells in growth medium, and can be detected with any luminometer; automated injectors are not required.
Host Cell Line: CHO-K1
Storage Stability: Immediately upon receipt, store vials in liquid nitrogen.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-1
References: 1. Delude, R.L., et.al. (1994) CD14-mediated Translocation of Nuclear Factor-kB Induced by Lipopolysaccharide Does Not Require Tyrosine Kinase Activity. J. Biol. Chem. 269: 222532. Railo, A., et.al. (2008) Wnt-11 signaling leads to down-regulation of the Wnt/beta-catenin, JNK/AP-1 and NF-kappaB pathways and promotes viability in the CHO-K1 cells. Exp Cell Res. 314: 2389-993. Murphy, S.H., et.al. (2011) Tumor suppressor protein (p)53, is a regulator of NF-κB repression by the glucocorticoid receptor. Proc. Natl. Acad. Sci. USA 108: 17117-17122
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