|HEK-Blue™ mTLR3 cells||Unit size||Cat. code||Docs||Qty||Price|
murine TLR3-expressing HEK 293 cells
3-7 x 10e6 cells
Murine TLR3 / NF-κB / SEAP reporter HEK293 cells
HEK-Blue™-mTLR3 cells were obtained by co-transfection of the murine 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™ mTLR3 cells is HEK-Blue™ Null1-k cells.
HEK-Blue™ mTLR3 and HEK-Blue™ Null1-k cells (control cell line) were stimulated with various TLR and NOD agonists: Pam3CSK4 (TLR2 agonist; 300 ng/ml), Poly(I:C) HMW (TLR3 agonist; 1 µg/ml), Poly(I:C) LMW (TLR3 agonist; 1 µg/ml), Poly(A:U) (TLR3 agonist; 10 µg/ml), LPS-EB ultrapure (TLR4 agonist; 10 µg/ml), FLA-ST (flagellin from S. typhimurium, a TLR5 agonist; 10 ng/ml), R848 (TLR7/8 agonist; 10 µg/ml), ODN 1826 (class B ODN specific for mTLR9; 10 µg/ml), Tri-DAP (NOD1 agonist; 3 µg/ml), MDP (NOD2 agonist; 10 µg/ml), and TNF-α (10 ng/ml). After 24h 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.
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™.
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- 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)
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