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HEK-Blue™ IL-18 Cells

HEK-Blue™ IL-18 cells Unit size Cat. code Docs Qty Price
3-7 x 10e6 cells
hkb-hmil18
+-
$1,142.00

IL-18 Sensor Cells

HEK-Blue™ IL-18 cells are designed to detect bioactive IL-18 by monitoring the activation of the NF-κB and AP-1 pathways. They were generated by stable transfection of HEK293 derived cells with the genes encoding IL-18R and IL-18RAP. In addition, the TNF-α and the IL-1β responses have been blocked. Therefore, HEK-Blue™ IL-18 cells respond specifically to IL-18. These cells express an NF-κB/AP-1-inducible SEAP reporter gene.

Binding of IL-18 to the heterodimeric IL-18 receptor on the surface of these cells triggers a signaling cascade leading to the activation NF-κB and the subsequent production of SEAP. Levels of SEAP in the supernatant can be easily determined with QUANTI-Blue™.


Response of HEK-Blue™ IL-18 cells to increasing concentrations of human and mouse recombinant IL-18. After a 24h incubation, the levels of SEAP were determined using QUANTI-Blue™, a SEAP detection reagent, and by reading the optical density (OD) at 630 nm.


Stimulation of HEK-Blue™ IL-18/IL-1β cells by recombinant human IL-18, IL-1β and TNF-α. After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ and reading the optical density (O.D.) at 655 nm.

Ligand EC50 Response Ratio
IL-18 2 +/- 1 ng/ml 55
 IL-1β 3 +/- 2 ng/ml 30

Note: 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.


HEK-Blue™ IL-18/IL-1β cells were stimulated with various human recombinant cytokines; IFNα (1000 IU/ml), IFNβ (1000 IU/ml), IFNγ (1000 IU/ml), IL-1β (100 ng/ml), IL-4 (100 ng/ml), IL-6 (100 ng/ml), IL-13 (100 ng/ml), IL-18 (100 ng/ml), TGF-β (10 ng/ml), and TNF-α (100 ng/ml). After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ and reading the O.D. at 655 nm.

Neutralization of IL-18 response
Neutralization of IL-18 response

Anti-hIL-18-IgA was incubated with 1 ng/ml of hIL-18 for 30 minutes prior to the addition of HEK-Blue™ IL-18/IL-1β cells, which were incubated with the antibody and cytokine for a further 24 h. Levels of SEAP in the supernatant were measured using QUANTI-Blue™ and reading the O.D. at 655 nm.


Anti-hIL-1β-IgA was incubated with 1 ng/ml of hIL-1β for 30 minutes prior to the addition of HEK-Blue™ IL-18/IL-1β cells, which were incubated with the antibody and cytokine for a further 24 h. Levels of SEAP in the supernatant were measured using QUANTI-Blue™ and reading the O.D. at 655 nm.

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Specifications

Antibiotic resistance: blasticidin, hygromycin B, Zeocin™

Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum (30 min at 56°C), 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Shipped on dry ice

Specificity: Detects human & murine IL-18

Detection range: 10 pg - 1 ng/ml for human IL-18, 3 ng - 100 ng/ml for mouse IL-18.

 

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Contents

• 1 vial containing 3-7 x 106 cells.

• 2 x 1 ml HEK-Blue™ Selection (250X concentrate); a solution containing several selection antibiotics.

• 1 ml Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria and fungi.

• 1 pouch of QUANTI-Blue™ (SEAP detection medium).

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Description

HEK-Blue IL18 pathway

Interleukin-18 (IL-18) is a pro-inflammatory cytokine that causes a wide variety of biological effects associated with infection, inflammation and autoimmune processes. It binds to an heterodimeric receptor consisting of IL-18R and IL-18 receptor accessory protein (IL18RAP) which then induces signaling pathways that involve MyD88, and TRAF-6 leading to the activation of NF-κB and AP-1.

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Citations

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