NF-κB reporter (Luc) - HEK293 Recombinant Cell line

Catalog #: 60650
Size: 2 vials
Species: Human
Nuclear factor-κB (NF-κB)/Rel proteins include NF-κB2 p52/p100, NF-κB1 p50/p105, c-Rel, RelA/p65, and RelB. These proteins function as dimeric transcription factors that control genes regulating a broad range of biological processes including innate and adaptive immunity, inflammation, stress responses, B cell development, and lymphoid organogenesis. In the classical (or canonical) pathway, NF-κB/Rel proteins are bound and inhibited by IκB proteins. Proinflammatory cytokines, LPS, growth factors, and antigen receptors activate an IKK complex (IKKβ, IKKα, and NEMO), which phosphorylates IκB proteins. Phosphorylation of IκB leads to its ubiquitination and proteasomal degradation, freeing NF-κB/Rel complexes. Active NF-κB/Rel complexes are further activated by phosphorylation and translocate to the nucleus where they induce target gene expression. In the alternative (or noncanonical) NF-κB pathway, NF-κB2 p100/ RelB complexes are inactive in the cytoplasm. Signaling through a subset of receptors, including LTβR, CD40, and BR3, activates the kinase NIK, which in turn activates IKKα complexes that phosphorylate C-terminal residues in NF-κB2 p100. Phosphorylation of NF-κB2 p100 leads to its ubiquitination and proteasomal processing to NF-κB2 p52, creating transcriptionally competent NF-κB p52/RelB complexes that translocate to the nucleus and induce target gene expression.
The NF-κB reporter (luc)-HEK293 cell line is designed for monitoring the nuclear factor Kappa B (NF-κB) signal transduction pathways. It contains a firefly luciferase gene driven by four copies of 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.
Materials Required But Not Provided:

• TNFα (Sigma # T0157-10UG)
• IL-1β (BPS Bioscience #90168)
• Assay Medium: Thaw Medium 1 (BPS Bioscience #60187)
• Growth Medium 1C (BPS Bioscience #79532)
• IKK-16 dihydrochloride (Sigma #SML1138): inhibitor of NF-κB activation
• 96-well tissue culture treated white clear-bottom assay plate (Corning #3610)
• One-Step luciferase assay system (BPS Bioscience #60690) or other luciferase reagents for measuring firefly luciferase activity
• Luminometer

NF-κB cell line, NFkappaB cell line, luciferase reporter, NF-kB cell line, NFkB cell line
Supplied As:
Each vial contains ~2 X 106 cells in 1 ml of 10% DMSO.
Biological Activity:
This cell line is validated for the response to TNFalpha and to treatment with NF-κB inhibitor, evodiamine.
Host Cell line:
HEK293 cells
Mycoplasma Testing:
The cell line has been screened using the PCR-based VenorGeM Mycoplasma Detection kit (Sigma Aldrich) to confirm the absence of Mycoplasma species.
Growth Media:

Thaw Medium 1 (BPS Bioscience Cat. #60187): MEM medium (Hyclone #SH30024.01) supplemented with 10% FBS, 1% non-essential amino acids (Hyclone #SH30238.01), 1 mM Na pyruvate (Hyclone #SH30239.01), 1% Penicillin/Streptomycin (Hyclone SV30010.01).

Growth Medium 1C (BPS Bioscience Cat. #79532): Thaw Medium 1 (BPS Bioscience #60187) and 50 μg/ml of Hygromycin B (Hyclone #SV30070.01).

Aqueous solution containing DMSO.
Instructions for use:
See data sheet for detailed culturing and assay protocol.
Storage / Stability:

Store in liquid nitrogen immediately upon receipt.

The NF-κB reporter cell line is designed for screening inhibitors of NF-κB and for monitoring NF-κB signaling pathway activity.

1. Pessara U, Koch N (1990). Mol Cell Biol.10(8):4146-4154.
2. Baeuerle PA (1998). Curr Biol. 8(1):R19-R22.
3. Takada Y, Kobayashi Y, Aggarwal BB (2005). J Biol Chem. 280(17):17203-17212.

Application Reference(s):
Synthesis of novel C5-curcuminoid-fatty acid conjugates and mechanistic investigation of their anticancer activity (2015)


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 for details. Publications using this cell line should reference BPS Bioscience, Inc., San Diego.

Avoid freeze/thaw cycles
Scientific Category:
Reporter Cell line
Product Type:
Cell line
Data shown is lot-specific. Contact us for specific information on other lots.

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