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IL-18 Reporter HEK 293 Cells

HEK-Blue™ IL-18 cells Unit size Cat. code Docs Qty Price
Human & Mouse IL-18 Reporter Cells
3-7 x 10e6 cells
hkb-hmil18
+-
$1,223.00

You may also need : Anti-hIL-18 | View more associated products

CONTENTS UPDATE NOTIFICATION
Please note that, for your convenience, these cells are now provided with QUANTI-Blue™ Solution.

HEK-Blue™ IL-18 Cells signaling pathway
HEK-Blue™ IL-18 Cells signaling pathway

IL-18 Reporter Cells

HEK-Blue™ IL-18 cells were engineered from the human embryonic kidney 293 (HEK293) cell line to detect bioactive interleukin-18 (IL-18) by monitoring the activation of the NF-κB and AP-1 pathways. These cells are useful to monitor IL-18 secretion in inflammasome activation studies. They can also be used for screening anti-IL-18 antibodies.

IL-18 is a pro-inflammatory cytokine that causes a wide variety of biological effects associated with infection, inflammation, and autoimmune processes [1, 2]. It is produced as a pro-protein which is proteolytically processed to its active form by caspase 1, an enzyme that is activated within the inflammasome multiprotein complex.

IL-18 binds to a heterodimeric receptor consisting of IL-18R and IL-18 receptor accessory protein (IL-18RAP). Upon binding, IL-18 activates NF-κB and AP-1 via signaling pathways that involve TRAF-6 [3, 4].

HEK-Blue™ IL-18 cells were generated by stable transfection of HEK293-derived cells with the genes encoding IL-18R and IL-18RAP. In addition, the responses to human TNF-α and IL-1β have been blocked. Therefore, HEK-Blue™ IL-18 cells respond specifically to IL-18. They are highly sensitive to human IL-18 and also respond to a lesser extent to murine IL-18.

HEK-Blue™ IL-18 cells express an NF-κB/AP-1-inducible secreted embryonic alkaline phosphatase (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 of NF-κB and the subsequent production of SEAP. Levels of SEAP in the supernatant can be easily determined with QUANTI-Blue™ Solution.

 

Features of HEK-Blue™ IL-18 cells:

  • Fully functional IL-18 signaling pathway
  • Do not respond to human TNF-α nor to IL-1β
  • Readily assessable SEAP reporter activity
  • Functionally tested and guaranteed mycoplasma-free

Applications of HEK-Blue™ IL-18 cells:

  • Detection of human IL-18 (10 pg - 1 ng/ml), and to a lesser extent murine IL-18 (3 -100 ng/ml)
  • Screening of anti-IL-18 antibodies

 

References:

1. Yasuda K. et al., 2019. Interleukin-18 in health and disease. Int J Mol Sci. 20(3).
2. Dinarello CA. et al., 1998. Overview of interleukin-18: more than an interferon-gamma inducing factor. J Leukoc Biol. 63(6):658-64.
3. Gracie JA. et al., 2003. Interleukin-18. J Leukoc Biol.3(2):213-24.
4. Kojima H. et al., 1998. Interleukin-18 activates the IRAK-TRAF6 pathway in mouse EL-4 cells. BBRC 244(1):183-6.

Figures


Cells were stimulated with increasing concentrations of recombinant human IL-18 (hIL-18) and murine  IL-18 (mIL-18). After overnight incubation, the NF-κB/AP-1 response was determined using QUANTI-Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 630 nm.

Cytokine response profile of HEK-Blue™ IL-18 cells
Cytokine response profile of HEK-Blue™ IL-18 cells

Response of HEK‑Blue™ IL‑18 cells to a panel of cytokines. Cells were stimulated with various human and murine recombinant cytokines: 1  ng/ml of hIL-18, mIL-18, 10  ng/ml of hIL-1β, hIL-33, hIFN-γ, hTNF-α, or 102 U/ml hIFN-α or hIFN-β. After overnight incubation, SEAP activity was assessed using QUANTI-Blue™ Solution. The optical density (OD) at 630 nm is shown as mean ± SD.

Human IL-18 signaling inhibition
Human IL-18 signaling inhibition

Dose‑dependent inhibition of HEK‑Blue™ IL‑18 cellular response using a neutralizing antibody against hIL‑18. The anti-hIL18 antibody was incubated with the cells for 30 minutes prior to the addition of hIL-18 (10  pg/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed   using QUANTI-Blue™ Solution. Data represent % of maximal reporter activity without the anti-hIL18 antibody.

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

Guaranteed mycoplasma-free

Specificity: Detects human & murine IL-18

Detection ranges:  

  • 10 pg - 1 ng/ml for human IL-18 
  • 3 ng - 100 ng/ml for mouse IL-18

 

These cells are covered by a Limited Use License (See Terms and Conditions).

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Contents

  • 1 vial containing 3-7 x 106 cells
  • 2 x 1 ml of HEK-Blue™ Selection (250x concentrate)
  • 1 ml of Normocin™ (50 mg/ml)
  • 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)

Shipped on dry ice (Europe, USA & Canada)

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Details

IL-18, formerly called interferon-γ (IFN-γ) inducing factor, is a pro-inflammatory cytokine that causes a wide variety of biological effects associated with infection, inflammation and autoimmune processes. More specifically, IL-18 induces IFN-γ production and contributes to T-helper 1 (Th1) cell polarization. IL-18 is produced by macrophages and other cells, as a pro-protein which is proteolytically processed to its active form by caspase 1, an enzyme that is activated within the inflammasome multiprotein complex.

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FAQ

Any questions about our cell lines ? Visit our frequently asked questions page

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Citations

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