The NF-κB reporter (Luc)-NIH/3T3 cell line is designed for monitoring nuclear factor Kappa B (NF-κB) signal transduction pathways. It contains a firefly luciferase gene driven by 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.

• Mouse TNFα (BioLegend #754402-10 μG)

• Assay medium: Thaw Medium 5 (BPS Bioscience, #60182) Or use o DMEM medium (Hyclone #SH30243.01) + 10% Calf Bovine Serum (Hyclone #SH3007203) + 1% Pen/Strep (Hyclone #SV30010.01)

• Growth Medium 5A (BPS Bioscience, #79534) 

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

• One-Step luciferase assay system (BPS Bioscience #60690)

• Luminometer

NF-kB reporter, NFkappaB reporter, luciferase reporter, NF-kB reporter, NFkB reporter, nih3t3

Each vial contains ~2 x 106 cells in 1 ml of 10% DMSO.

NF-κB reporter (Luc)-NIH/3T3

The cell line has been screened using the PCR-based Venor^®GeM Mycoplasma Detection kit (Sigma-Aldrich, #MP0025) to confirm the absence of Mycoplasma species.

Thaw Medium 5 (BPS Cat. # 60182): DMEM medium (Hyclone #SH30243.01) supplemented with 10% Calf Serum (CS) (Hyclone # SH30072.03), not FBS, 1% Penicillin/Streptomycin (Hyclone # SV30010)

Growth Medium 5A (BPS Cat. #79534): Thaw Medium 5 (BPS Cat. #60182) plus 600 μg/ml of Geneticin (Life Technologies #11811031).

See data sheet for detailed culturing and assay protocol.

Immediately upon receipt, store in liquid nitrogen.

• Monitor NF-κB signaling pathway activity.

• Screen for activators or inhibitors of NF-κB signaling pathway.

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.

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 the 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 Bioscience 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.

Avoid freeze/thaw cycles

Cell Signaling Pathway

Cell line