HEK-Blue™ TNF-α Cells

September 12^th 2019 - CONTENTS UPDATE NOTIFICATION
Please note that, for your convenience, these cells are now provided with QUANTI-Blue™ Solution.

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.

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

Specifications

Antibiotic resistance: Puromycin, Zeocin™

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™

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)

Contents

• 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)

Description

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.