|HEK-Blue™ hNOD1 cells||Unit size||Cat. code||Docs||Qty||Price|
human NOD1-expressing HEK 293 cells
3-7 x 10e6 cells
Human NOD1-expressing HEK 293 cells
HEK-Blue™-hNOD1 cells were obtained by co-transfection of the human NOD1 gene and an optimized secreted embryonic alkaline phosphatase (SEAP) reporter gene 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. Stimulation with a NOD1 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™ hNOD1 cells is HEK-Blue™ Null1 cells.
HEK-Blue™ hNOD1 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 reading the OD at 655 nm.
HEK-Blue™ hNOD1 and HEK-Blue™ Null1 (control) cells were incubated in HEK-Blue™ Detection medium and stimulated with 100 ng/ml C12-iE-DAP (NOD1), 1 µg/ml M-TriDAP (NOD1/2), 1 µg/ml Tri-DAP (NOD1), or 1 µg/ml iE-DAP (NOD1). After 24h incubation, the levels of NF-κB-induced SEAP were determined by reading the OD at 655 nm.
HEK-Blue™ hNOD1 cells were incubated in HEK-Blue™ Detection medium and stimulated with increasing concentrations of NOD1 agonists. After 24h incubation, the levels of NF-κB-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.
|C12-iE-DAP||5 +/- 2 ng/ml||10|
|Tri-DAP||700 +/- 100 ng/ml||6|
|iE-DAP||6.3 +/- 0.5 µg/ml||5|
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™
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- 1 vial containing 3-7 x 106 cells
- 100 μl Blasticidin (10 mg/ml)
- 100 μl Zeocin™ (100 mg/ml)
- 1 ml Normocin™ (50 mg/ml)
- 1 pouch of HEK-Blue™ Detection (cell culture medium for real-time detection of SEAP)
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