HEK-Blue™ mTLR9

HEK-Blue™ mTLR9 cells Unit size Cat. code Docs Qty Price
murine TLR9-expressing HEK 293 cells
3-7 x 10e6 cells

HEK-Blue™-mTLR9 cells were obtained by co-transfection of the murine TLR9 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 TLR9 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™ mTLR9 cells is HEK-Blue™ Null1 cells.

HEK-Blue™ mTLR9 and HEK-Blue™ Null1 cells (control cell line) were stimulated with various TLR and NOD agonists: Pam3CSK4 (TLR2 agonist; 100 ng/ml), Poly(I:C) (TLR3 agonist; 1 µg/ml), LPS-EB ultrapure (TLR4 agonist; 100 ng/ml), FLA-ST (flagellin from S. typhimurium, a TLR5 agonist; 100 ng/ml), R848 (TLR7/8 agonist; 1 µg/ml), ODN 1668 (class B ODN specific for mTLR9; 10 µg/ml), ODN 1826 (class B ODN specific for mTLR9; 10 µg/ml), ODN 2006 (class B ODN with a preference for human TLR9; 10 µg/ml), Tri-DAP (NOD1 agonist; 1 µg/ml), MDP (NOD2 agonist; 1 µg/ml), and TNF-α (10 ng/ml). After a 24h incubation, NF‑kB‑induced SEAP activity was assessed using HEK‑Blue™ Detection and reading the optical density (OD) at 655 nm. TNF-α has been included as a positive control. Non‑induced cells (NI) have been included as negative controls.

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Antibiotic resistance: blasticidin, 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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Shipped on dry ice

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