|HEK-Blue™ hTLR9 cells||Unit size||Cat. code||Docs||Qty||Price|
human TLR9-expressing HEK 293 cells
3-7 x 10e6 cells
SEAP Reporter 293 cells expressing the human TLR9 gene
HEK-Blue™ hTLR9 cells were obtained by co-transfection of the human TLR9 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 TLR9 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™ hTLR9 cells is HEK-Blue™ Null1 cells.
HEK-Blue™ hTLR9 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-kB-induced SEAP activity was assessed using QUANTI-Blue™ and reading the OD at 655 nm.
HEK-Blue™ hTLR9 and HEK-Blue™ Null1 (control) cells were incubated in HEK-Blue™ Detection medium and stimulated with 30 µg/ml ODN2216 (human specific, type A), 0.3 µg/ml ODN2006 (human specific, type B), 30 µg/ml ODN2395 (human specific, type C), 30 µg/ml ODN1826 (mouse specific, type B), or 5 µg/ml E.coli ssDNA/LyoVec™. After 18h incubation, NF-kB-induced SEAP activity was assessed using QUANTI-Blue™ and reading the OD at 655 nm.
HEK-Blue™ hTLR9 cells were stimulated with increasing concentrations of TLR9 agonists. In the top graph, the response to ODN2006 (human specific, type B) is shown. In the bottom graph, the response to ODN2216 (human specific, type A) and ODN2395 (human/mouse, type C) is presented. After 18h incubation, NF-κB-induced SEAP activity was assessed using QUANTI-Blue™. 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.
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.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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