Human TLR2 Reporter HEK293 Cells (NF-κB)

HEK-Blue™ hTLR2 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 (3 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™ hTLR2 and HEK-Blue™ Null1 (control) cells were incubated in HEK-Blue™ Detection medium and stimulated with 0.1 ng/ml FSL-1 (TLR2/6), 10 ng/ml Pam3CSK4 (TLR1/2), 107 cells/ml HKLM, 1 µg/ml LTA-SA, 10 ng/ml LPS-PG, or 1 µg/ml PGN-BS. After 24h incubation, the levels of NF-κB-induced SEAP were determined by reading the OD at 655 nm.

Anti-hTLR1 IgG         IC50 400 +/-100 ng/ml
Anti-hTLR2 IgA2       IC50 350 +/-50 ng/ml

HEK-Blue™ TLR2 cells were pre-incubated for 1h with increasing concentrations of anti-hTLR2-IgA, anti-hTLR6-IgG or anti-hTLR1-IgG then stimulated with 0.5 ng/ml of Pam3CSK4 for 18h. NF-kB-induced SEAP activity was assessed using QUANTI-Blue™ by reading the OD at 655 nm.

Anti-hTLR2 IgA2       IC50 300 +/-100 ng/ml
Anti-hTLR6 IgG         IC50 60 +/-20 ng/ml

HEK-Blue™ TLR2 cells were pre-incubated for 1h with increasing concentrations of anti-hTLR2-IgA, anti-hTLR6-IgG or anti-hTLR1-IgG then stimulated with 0.1 ng/ml of FSL-1 for 18h. NF-kB-induced SEAP activity was assessed using QUANTI-Blue™ by reading the OD at 655 nm.