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.

• TNFα (BPS Bioscience, #90245)

• IL-1β (BPS Bioscience, #90168)

• Growth Medium 1C (BPS Bioscience, #79532)

• Evodiamine (Abcam, #142427): inhibitor of NF-κB activation

• 96-well tissue culture treated white clear-bottom assay plate (Corning, #3610)

• Luminometer

Thaw Medium 1 (BPS Bioscience, #60187): Medium optimized for culturing HEK293 cells.

Growth Medium 1C (BPS Bioscience, #79532): Thaw Medium 1 plus 50 µg/ml of Hygromycin B (Hyclone, #SV30070.01).

• Screen inhibitors of NFκB

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 NFκB-regulated antiapoptotic and metastatic gene expression, up-regulating apoptosis, and inhibiting invasion. J. Biol. Chem. 280(17):17203-17212

License Disclosure

Purchase of this cell line grants you with a 10-year license to use this cell line in your immediate laboratory, for research use only. This license does not permit you to share, distribute, sell, sublicense, or otherwise make the cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit use of this cell line in humans or for therapeutic or drug use. The license does not permit modification of the cell line in any way. Inappropriate use or distribution of this cell line will result in revocation of the license and result in an immediate cease of sales and distribution of BPS products to your laboratory. BPS does not warrant the suitability of the cell line for any particular use, and does not accept any liability in connection with the handling or use of the cell line. Modifications of this cell line, transfer to another facility, or commercial use of the cells may require a separate license and additional fees; contact sales@bpsbioscience.com for details. Publications using this cell line should reference BPS Bioscience, Inc., San Diego.