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HEK-Blue™ hTLR2

HEK-Blue™ hTLR2 cells Unit size Cat. code Docs Qty Price
human TLR2-expressing HEK 293 cells
3-7 x 10e6 cells
hkb-htlr2
+-
$1,142.00

HEK-Blue™-hTLR2 cells were obtained by co-transfection of the human TLR2 and SEAP (secreted embryonic alkaline phosphatase) genes 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.

Additionally, the CD14 co-receptor gene was transfected into these cells to enhance the TLR2 response.

Stimulation with a TLR2 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™ hTLR2 cells is HEK-Blue™ Null1 cells.

Response of HEK-Blue™ hTLR2 cells to TLR and NOD agonists
Response of HEK-Blue™ hTLR2 cells to TLR and NOD agonists

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 Ultrapure 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.

TLR2 agonists dose response
TLR2 agonists dose response

HEK-Blue™ hTLR2 cells were stimulated with increasing concentrations of TLR2 agonists. After 24h incubation, NF-kB-induced SEAP activity was assessed using HEK-Blue™ Detection 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.

Ligand EC50 Response Ratio
 FSL-1 0.6 +/- 0.1 ng/ml 15
 Pam3CSK4 0.1 +/- 0.1 ng/ml  14
Neutralization of TLR1/2 dose response
Neutralization of TLR1/2 dose response

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.

Neutralization of TLR2/6 dose response
Neutralization of TLR2/6 dose response

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.

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Specifications

Antibiotic resistance: blasticidin, hygromycin, Zeocin™

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

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Contents

Shipped on dry ice

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