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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)-HEK293 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 1) that has been optimized for use with HEK293 cells. Thaw Medium 1 includes 10% fetal bovine serum, non-essential amino acids, sodium pyruvate, 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: HEK293
Storage Stability: Immediately upon receipt, store vials in liquid nitrogen.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: 1. Pessara U, Koch N (1990) Tumor necrosis factor alpha regulates expression of the major histocompatibility complex class II-associated invariant chain by binding of an NF-κB-like factor to a promoter element. Mol. Cell. Biol. 10(8):4146-4154.2. Baeuerle PA (1998) Pro-inflammatory signaling: last pieces in the NF-κB puzzle? Curr. Biol. 8(1):R19-R22.3. Takada Y, Kobayashi Y, Aggarwal BB (2005) Evodiamine abolishes constitutive and inducible NF-κB activation by inhibiting IκBα kinase activation, thereby suppressing NF-κB-regulated antiapoptotic and metastatic gene expression, up-regulating apoptosis, and inhibiting invasion. J. Biol. Chem. 280(17):17203-17212
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