IL-18 Reporter HEK 293 Cells
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Human & Mouse IL-18 Reporter Cells
3-7 x 10e6 cells
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
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.
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.
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.
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.
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™
Specificity: Detects human & murine IL-18
- 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).Back to the top
- 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)Back to the top
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.Back to the top