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Human TLR5 Reporter HEK293 Cells (NF-κB)

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HEK-Blue™ hTLR5 cells

Human TLR5 expressing HEK293 reporter cells (NF-κB pathway)

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3-7 x 10e6 cells

hkb-htlr5
+-
$1,457

Notification:  This product is for internal research use only. Additional rights may be available. Please visit InvivoGen’s Terms and Conditions.

NF-κB–SEAP reporter HEK293 cells expressing human TLR5

HEK-Blue™-hTLR5 cells were obtained by co-transfection of the human TLR5 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 TLR5 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™ hTLR5 cells is HEK-Blue™ Null1 cells.

Figures

Response of HEK-Blue™ hTLR5 cells to TLR and NOD agonists
Response of HEK-Blue™ hTLR5 cells to TLR and NOD agonists

HEK-Blue™ hTLR5 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), recFLA-ST (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™ by reading the OD at 655 nm.

Response of HEK-Blue™ hTLR5  to TLR5 agonists
Response of HEK-Blue™ hTLR5  to TLR5 agonists

HEK-Blue™ hTLR5 and HEK-Blue™ Null1 (control) cells were incubated in HEK-Blue™ Detection medium and stimulated with 100 ng/ml FLA-BS, 10 ng/ml FLA-ST Ultrapure, or 10 ng/ml recFLA-ST. After 24h incubation, the levels of NF-κB-induced SEAP were determined by reading the OD at 655 nm.

TLR5 agonists dose response
TLR5 agonists dose response

HEK-Blue™ hTLR5 cells were stimulated with increasing concentrations of TLR5 agonists. After 24h incubation, NF-κB-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 Origin EC50 Response Ratio
 FLA-BS B.subtilis 15 +/- 5 ng/ml 29
 FLA-ST S.typhimurium 6 +/- 3 ng/ml 23
Neutralization of TLR5 dose response
Neutralization of TLR5 dose response

HEK-Blue™ TLR5 cells were pre-incubated for 1h with increasing concentrations of anti-hTLR5 IgA2, then stimulated with 5 ng/ml of FLA-ST Ultrapure 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, Zeocin®.

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.

 

This product is covered by a Limited Use License (See Terms and Conditions).

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Contents

Shipped on dry ice (Europe, USA, Canada, and some areas in Asia)

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