CD40/NF-κB Luciferase Reporter HEK293 Cell Line
Recombinant HEK293 cell line expressing the full length human CD40 (also known as Tumor necrosis factor receptor superfamily member 5; TNFRSF5). Expression is confirmed by flow cytometry and western blot. An NF-ĸB luciferase reporter construct is stably integrated into the genome. Thus, the firefly luciferase gene is controlled by 4 copies of an NF-kB response element located upstream of a TATA promoter. Following activation by a CD40 agonist, NF-ĸB transcription factors bind to the DNA response elements to induce transcription of the luciferase gene. Activation of CD40 is quantified by measuring luciferase activity.
Illustration of CD40/NF-κB Luciferase Reporter HEK293 Cell Line
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This product has been cited 2 times.
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Media Required for Cell Culture
Name | Ordering Information |
Thaw Medium 1 | BPS Bioscience #60187 |
Growth Medium 1A | BPS Bioscience #79528 |
Materials Used in the Cellular Assay
Name | Ordering Information |
human TNFα | R&D Systems #210-TA |
CD40L | BPS Bioscience #71191 |
CD40L antibody | BioLegend #310827 |
ONE-Step™ Luciferase Assay System | BPS Bioscience #60690 |
White clear-bottom 96-well cell culture plate | |
Luminometer |
The cell line has been screened to confirm the absence of Mycoplasma species.
CD40, a member of the TNF receptor superfamily, is expressed in B lymphocytes, antigen presenting cells (APCs) such as monocytes, basophils, and dendritic cells, and in non-immune cells such as endothelial and epithelial cells. The protein is overexpressed in several types of carcinomas. Interaction of CD40 with CD40 ligands (CD40L, CD154) on CD4+ T helper lymphocytes triggers the expression of intercellular adhesion molecule (ICAM) and other pro-inflammatory cytokines. CD40:CD40L signaling increases the activation of antigen-specific T cells. CD40 also activates NF-κB-dependent signaling in response to lipopolysaccharide (LPS) found on Gram negative bacterial pathogens. Furthermore, agonistic CD40 monoclonal antibodies activate antigen presenting cells (APC) and promote anti-tumor T-cell responses in addition to fostering cytotoxic myeloid cells with the potential to control cancer in the absence of T cell immunity. CD40 is considered a potential target for cancer immunotherapy.
1. Li G et.al. (2013) Human Genetics in Rheumatoid Arthritis Guides a High-Throughput Drug Screen of the CD40 Signaling Pathway. PLoS Genet. 9: e1003487.
2. Pontrelli P et.al. (2006) CD40L Proinflammatory and Profibrotic Effects on Proximal Tubular Epithelial Cells: Role of NF-κB and Lyn. J. Amer. Soc. Neph. 17: 627.
3. Lavorgna A et.al. (2014) A Critical Role for IL-17RB Signaling in HTLV-1 Tax-Induced NF-κB Activation and T-Cell Transformation. PLoS Path. 10: e1004418.
4. Moschonas A et.al. (2012) CD40 Stimulates a “Feed-Forward” NF-ĸB- Driven Molecular Pathway That Regulates IFN-β Expression in Carcinoma Cells. J. Immunol. 188: 5521.
Application References:
Intein mediated high throughput screening for bispecific antibodies