|HEK-Dual™ hTLR3 (NF/IL8)||Unit size||Cat. code||Docs||Qty||Price|
Double readout TLR cells
3-7 x 10e6 cells
Human TLR3 (NF-κB-SEAP / KI-[IL-8]Lucia) dual-reporter HEK293 cells
HEK-Dual™ hTLR3 (NF/IL8) cells were generated from HEK-Dual™ Null cells by stable transfection of the human TLR3 (hTLR3) gene. Due to the knockout of TLR5, these cells enable the study of hTLR3 signaling without interference from other TLRs.
They respond to very low concentrations of TLR3 agonists such as poly(I:C) a synthetic analog of double-stranded RNA. They do not respond to other TLR agonists or to the cytokine TNF-α (see validation sheet).
1. Ohta K. et al., 2014. TLR-mediated interleukin-8 production by human submandibular gland epithelial cells. Mol Med Rep. 10(5):2377-82.
2. Roebuck KA. et al., 1999. Regulation of interleukin-8 gene expression. J Interferon Cytokine Res. 19(5):429-38.
Detection of the IL-8 response using QUANTI-Luc™. HEK-Dual™ hTLR3 (NF/IL8) cells were stimulated with TLR3 agonists: Poly(I:C) HMW (100 ng/ml), Poly(I:C) LMW (100 ng/ml) and Poly(A:U) (100 ng/ml). After 24 hour incubation, activation of the IL‐8 promoter was determined by measuring the relative light units (RLUs) in a luminometer using QUANTI-Luc™, a Lucia luciferase detection reagent. The activation of the IL‐8 promoter is expressed as fold increase relative to untreated cells which was calculated by dividing the RLUs for the treated cells by the RLUs for the untreated cells.
Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) fetal bovine serum, 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™.
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- -1 vial containing 3-7 x 106 cells.
- 100 μl Hygromycin B Gold (100 mg/ml).
- 100 μl Zeocin™ (100mg/ml).
- 1 ml Normocin™ (50 mg/ml).
- 1 pouch of QUANTI-Blue™ (SEAP detection medium).
- 1 pouch of QUANTI-Luc™ (Lucia luciferase detection medium).
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EC50 values calculated for the NF-kB response using QUANTI-Blue™. HEK-Dual™ hTLR3 (NF/IL8) and HEK-Blue™ hTLR3 cells were stimulated for 24 hours with various TLR3 agonists: Poly(I:C) HMW, Poly(I:C) LMW and Poly(A:U).
|Cell line||EC50 for Poly(I:C) HMW (ng/ml)||EC50 for Poly(I:C) HMW (ng/ml)||EC50 for Poly(A:U) (ng/ml)|
|HEK-Dual™ hTLR3 (NF/IL8)||28.9 ± 17.7||59.4 ± 35.3||53.6 ± 22.9|
|HEK‑Blue™ hTLR3||77.6 ± 29.0||163.8 ± 99.9||218.5 ± 82.7|