HEK-Blue™ hTLR3

Human TLR3 / NF-κB–SEAP reporter HEK293 cells

HEK-Blue™-hTLR3 cells were obtained by co-transfection of the human TLR3 gene and an inducible SEAP (secreted embryonic alkaline phosphatase) reporter gene into HEK293 cells.

The SEAP gene is placed under the control of the IFN-β minimal promoter fused to five NF-κB and AP-1-binding sites. Stimulation with a TLR3 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™ hTLR3 cells is HEK-Blue™ Null1 cells.

Figures

HEK-Blue™ hTLR3 cells were stimulated with various TLR and NOD agonists: Pam3CSK4 (100 ng/ml), Poly(I:C)-HMW (50 ng/ml), LPS-EB ultrapure (100 ng/ml), recombinant flagellin from S. typhimurium (10 ng/ml), CL264 (1 µg/ml), CL097 (1 µg/ml), ssRNA40/LyoVec™ (5 µg/ml), ODN 2006 (10 µg/ml), C12-iE-DAP (100 ng/ml), L18-MDP (100 ng/ml), and TNF-α (100 ng/ml). After 18h incubation (24h incubation for CL264, C12-iE-DAP and L18-MDP ligands), NF-κB-induced SEAP activity was assessed using QUANTI-Blue™ and by reading the OD at 655 nm.

HEK-Blue™ hTLR3 and HEK-Blue™ Null1 (control) cells were stimulated with poly(I:C)-HMW or poly(I:C)-LMW. All agonists were used at a concentration of 1 µg/ml. After 18h incubation, NF-κB-induced SEAP activity was assessed using QUANTI-Blue™ by reading the OD at 655 nm.

HEK-Blue™ hTLR3 cells were stimulated with increasing concentrations of TLR3 agonists. After 18h incubation, NF-κB-induced SEAP activity was assessed using QUANTI-Blue™ and by reading the OD at 655 nm. The response ratio was calculated by dividing the OD at 655 nm for the treated cells by the OD at 655 nm for the untreated cells.

 

Ligand	EC50	Response Ratio
Poly(I:C) HMW	20 +/- 5 ng/ml	19
Poly(I:C) LMW	40 +/- 5 ng/ml	18

Specifications

Antibiotic resistance: blasticidin, 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, 100 μg/ml Normocin™.

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
• 100 μl Blasticidin (10 mg/ml)
• 100 μl Zeocin® (100 mg/ml)
• 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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