HEK-Blue™ mTLR4

Murine TLR4 / NF-κB–SEAP reporter HEK293 cells

HEK-Blue™-mTLR4 cells were obtained by co-transfection of the murine TLR4, MD-2 and CD14 co-receptor genes, and an inducible SEAP (secreted embryonic alkaline phosphatase) reporter gene into HEK293 cells.

The SEAP reporter gene is placed under the control of an IFN-β minimal promoter fused to five NF-κB and AP-1-binding sites. Stimulation with a TLR4 ligand activates NF-κB and AP-1 which induce the production of SEAP.

Levels of SEAP can be easily determined with HEK-Blue™ Detection, a cell culture medium that allows for real-time detection of SEAP.

The parental cell line of HEK-Blue™ mTLR4 cells is HEK-Blue™ Null1-v cells.

Figures

HEK-Blue™ mTLR4 and HEK-Blue™ Null1-v cells (control cell line) were stimulated with various TLR and NOD agonists: Pam3CSK4 (TLR2 agonist; 100 ng/ml), Poly(I:C) (TLR3 agonist; 1 µg/ml), LPS-EB ultrapure (TLR4 agonist; 100 ng/ml), LPS-EB ultrapure (TLR4 agonist; 100 ng/ml), MPLA (TLR4 agonist; 100 ng/ml), FLA-ST (flagellin from S. typhimurium, a TLR5 agonist; 100 ng/ml), R848 (TLR7/8 agonist; 1 µg/ml), ODN 1826 (class B ODN; 10 µg/ml), Tri-DAP (NOD1 agonist; 1 µg/ml), MDP (NOD2 agonist; 1 µg/ml), and TNF-α (10 ng/ml). After a 24 hour-incubation, NF‑kB‑induced SEAP activity was assessed using HEK‑Blue™ Detection and reading the optical density (OD) at 655 nm. TNF-α has been included as a positive control. Non‑induced cells (NI) have been included as negative controls.

Specifications

Antibiotic resistance: blasticidin, hygromycin, Zeocin®.

Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin.

Guaranteed mycoplasma-free.

This product is covered by a Limited Use License (See Terms and Conditions).

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 2 ml 250X HEK-Blue Selection
• 1 ml Normocin™ (50 mg/ml)
• 1 pouch of HEK-Blue™ Detection (cell culture medium for real-time detection of SEAP)

Shipped on dry ice (Europe, USA, Canada and some areas in Asia)

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