HEK-Blue™ hNOD2

Human NOD2 / NF-kB / SEAP reporter HEK293 cells

HEK-Blue™-hNOD2 cells were obtained by co-transfection of the human NOD2 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 IL-12 p40 minimal promoter fused to five NF-κB and AP-1 binding sites. Stimulation with a NOD2 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™ hNOD2 cells is HEK-Blue™ Null2 cells.

Figures

HEK-Blue™ hNOD2 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™ hNOD2 and HEK-Blue™ Null2 (control) cells were incubated in HEK-Blue™ Detection medium and stimulated with 10 ng/ml L18-MDP (NOD2), 100 ng/ml MDP (NOD2), 100 ng/ml M-TriDAP (NOD1/2), 100 ng/ml murabutide (NOD2) or 100 ng/ml N-Glycolyl-MDP (NOD2). After 24h incubation, the levels of NF-κB-induced SEAP were determined by reading the OD at 655 nm.

HEK-Blue™ hNOD2 cells were incubated in HEK-Blue™ Detection medium and stimulated with increasing concentrations of NOD2 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.

Ligand	EC50	Response Ratio
L18-MDP	0.15 +/- 0.1 ng/ml	19
MDP	35 +/- 5 ng/ml	17
Murabutide	100 +/- 20 ng/ml	8

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, 100μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free.

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

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml of Blasticidin (10 mg/ml)
• 1 ml of Zeocin® (100 mg/ml)
• 1 ml of Normocin™ (50 mg/ml)
• 1 pouch of HEK-Blue™ Detection (cell culture medium for real-time detection of SEAP)

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

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