Human TLR4 Reporter HEK293 Cells (NF-κB)
|HEK-Blue™ hTLR4 cells||Unit size||Cat. code||Docs||Qty||Price|
Human TLR4 expressing HEK293 reporter cells (NF-κB pathway)
3-7 x 10e6 cells
NF-κB–SEAP reporter HEK293 cells expressing human TLR4
HEK-Blue™-hTLR4 cells were obtained by co-transfection of the human TLR4, MD-2 and CD14 co-receptor genes, and an inducible SEAP (secreted embryonic alkaline phosphatase) reporter gene into HEK293 cells.
The SEAP reporter gene is placed under the control of an IL-12 p40 minimal promoter fused to five NF-κB and AP-1-binding sites.
Stimulation with a TLR4 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™ hTLR4 cells is HEK-Blue™ Null2 cells.
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.
|LPS-EB up||2 +/- 1 ng/ml||13|
|LPS-EK||0.07 +/- 0.01 ng/ml||16|
|MPLAs||10 +/- 3 ng/ml||12|
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.
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™
This product is covered by a Limited Use License (See Terms and Conditions).Back to the top
1 vial containing 3-7 x 106 cells
- 2 ml 250X HEK-Blue™ Selection
- 1 ml Normocin™ (50 mg/ml)
- 1 pouch of HEK-Blue™ Detection (cell culture medium for real-time detection of SEAP)
Shipped on dry ice. (Europe, USA, Canada and some areas in Asia)Back to the top