HEK-Blue™ TNF-α Cells

HEK-Blue™ TNF-α cells Unit size Cat. code Docs Qty Price
Human TNF-α SEAP reporter HEK293 cells
3-7 x 10e6 cells

Human and murine TNF-α Sensor Cells

HEK-Blue™ TNF-α cells enable the detection of bioactive human and murine TNF-α by monitoring the activation of the NF-κB pathway.
These cells were generated by stable transfection of HEK293 cells with a SEAP reporter gene under the control of the IFN-β minimal promoter fused to five NF-κB (and five AP-1) binding sites. They were further rendered unresponsive to IL-1β by knocking-out the MyD88 gene.
Stimulation of HEK-Blue™ TNF-α cells with TNF-α triggers the activation of the NF-κB-inducible promoter and the production of SEAP. Levels of SEAP in the supernatant can be easily determined using QUANTI-Blue™, a reagent that turns purple/blue in the presence of SEAP, by reading the OD at 620-655 nm.

HEK-Blue™ TNF-α cells are resistant to Zeocin™ and puromycin.

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Antibiotic resistance: Zeocin™, puromycin

Growth medium: DMEM, 4.5 g/l glucose, 10% fetal bovine serum, 2 mM L-glutamine, 100 µg/ml Normocin™

Shipped on dry ice

Quality Control
Reporter activity is validated by stimulating the cells with human and murine TNF-α.
The cells are guaranteed mycoplasma-free.

Detects human and murine TNF-α
• hTNF-α EC50: 0.01 ng/ml (in medium) or 0.7 ng/ml (in water)
• mTNF-α EC50:
0.1 ng/ml (in medium) or 3 ng/ml (in water)

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1 vial of HEK-Blue™ TNF-α cells (3-7 x 10e6 cells) in freezing medium.
IMPORTANT: Cells are shipped frozen. If cells are not frozen upon arrival, contact InvivoGen immediately.
• 100 μl Zeocin™ (100 mg/ml).
• 100 μl puromycin (10 mg/ml).
• 1 ml Normocin™ (50 mg/ml).
• 1 pouch QUANTI-Blue™ (SEAP detection reagent).

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HEK-Blue TNF-alpha pathway

Tumor necrosis factor-alpha (TNF-α) is a pleiotropic inflammatory cytokine produced by several types of cells, predominantly activated macrophages.
TNF-α plays an important role in the immune response to microbial invasions and in the necrosis of specific tumors. TNF-α binds two receptors TNFR1 and TNFR2 inducing a signaling that involves TRADD, TRAF2 and RIP, and leads to the activation of the NF-κB and the MAPK pathways.
Interleukin 1 beta (IL-1β) is another inflammatory cytokine that triggers these pathways following the binding to its receptor IL-1RI and the recruitment of MyD88.

Both TNF-α and IL-1β receptors are expressed in HEK293 cells. HEK-Blue™ TNF-α Cells are rendered unresponsive to IL-1β by knocking-out the MyD88 gene.

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