HEK-Blue™ TNF-α Cells
|HEK-Blue™ TNF-α cells||Unit size||Cat. code||Docs||Qty||Price|
Human TNF-α SEAP Reporter Cells
3-7 x 10e6 cells
Human and murine TNF-α Reporter 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.Back to the top
Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 µg/ml streptomycin, 100 µg/ml Normocin™
- 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)
- 1 vial of HEK-Blue™ TNF-α cells (3-7 x 10e6 cells)
- 1 ml of Puromycin (10 mg/ml)
- 1 ml of Zeocin™ (100 mg/ml)
- 1 ml of Normocin™ (50 mg/ml)
- 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)
Shipped on dry ice (Europe, USA & Canada)Back to the top
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.Back to the top