|HEK-Blue™ hTLR7 cells||Unit size||Cat. code||Docs||Qty||Price|
human TLR7-expressing HEK 293 cells
3-7 x 10e6 cells
HEK-Blue™-hTLR7 cells were obtained by co-transfection of the human TLR7 gene and an inducible SEAP (secreted embryonic alkaline phosphatase) reporter gene into HEK293 cells.
The SEAP reporter 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 TLR7 ligand activates NF-κB and AP-1 which induces 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™ hTLR7 cells is HEK-Blue™ Null1-k cells.
HEK-Blue™ hTLR7 cells were stimulated with various TLR and NOD agonists: Pam3CSK4 (100 ng/ml), Poly(I:C) (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 reading the OD at 655 nm.
HEK-Blue™ hTLR7 and HEK-Blue™ Null1-k (control) cells were incubated in HEK-Blue™ Detection medium and stimulated with CL264 (TLR7), R848 (TLR7/8), Gardiquimod™ (TLR7), CL097 (TLR7/8), Imiquimod (TLR7), or CL075 (TLR7/8). All agonists were used at a concentration of 1 µg/ml. After 24h incubation, the levels of NF-κB-induced SEAP were determined by reading the OD at 655 nm.
HEK-Blue™ hTLR7 cells were incubated in HEK-Blue™ Detection medium and stimulated with increasing concentrations of TLR7 and TLR7/8 agonists. After 24h incubation, the levels of NF-κB-induced SEAP were determined 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.
|CL264||263 +/- 65 ng/ml||22|
|R848||42 +/- 22 ng/ml||22|
Growth medium: DMEM, 4.5 g/l glucose, 2-4 mM L-glutamine, 10% (v/v) fetal bovine serum, 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™.
Guaranteed mycoplasma-free.Back to the top
- 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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