Human TLR4 Reporter HEK293 Cells (NF-κB)

HEK-Blue™ hTLR4 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™ hTLR4 and HEK-Blue™ Null2 (control) cells were incubated in HEK-Blue™ Detection medium and stimulated with 10 ng/ml LPS-EB ultrapure, 10 ng/ml LPS-EK ultrapure, 100 ng/ml MPLA or 100 ng/ml MPLAs. After 24h incubation, the levels of NF-κB-induced SEAP were determined by reading the OD at 655 nm.

HEK-Blue™ hTLR4 cells were incubated in HEK-Blue™ Detection medium and stimulated with increasing concentrations of TLR4 agonists. After 24h incubation, the levels of NF-kB-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.

HEK-Blue™ TLR4 cells were pre-incubated for 1h with increasing concentrations of anti-hTLR4 IgA2, then stimulated with 5 ng/ml of LPS-EB Ultrapure for 18h. NF-kB-induced SEAP activity was assessed using QUANTI-Blue™ by reading the OD at 655 nm.