IL-1α & IL-1β Reporter HEK 293 Cells
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HEK 293 reporter cells for human and murine IL-1α & IL-1β cytokines
3-7 x 10e6 cells
Human & Murine IL-1 Reporter Cells
HEK-Blue™ IL-1R cells were designed to detect bioactive human and murine interleukin-1 cytokines (IL‑1α and IL-1β) in various biological samples (cell culture supernatant and serum) by monitoring the activation of the NF-κB and AP-1 pathways. Additionally, these cells detect bioactive IL-1β from cynomolgus monkeys, dogs, and rats
IL‑1 is a secreted pro‑inflammatory cytokine that plays a critical role in immune responses and inflammation . It exists in two distinct isoforms, IL-1α and IL-1β, which are produced as pro-proteins by activated macrophages. IL-1β is cleaved by caspase-1  while IL-1α is cleaved by calcium‑activated calpain or caspase-5 (or its murine ortholog caspase-11) . In contrast to pro‑IL‑1β, pro-IL-1α is active, although cleavage dramatically enhances its bioactivity. IL-1β and IL-1α bind to the same receptor, IL-1R1, triggering the formation of the IL-1R1/IL-1R3/MyD88 complex. This induces signaling leading to the activation of the transcription factors NK-κB and AP-1 with the subsequent inflammatory response .
HEK-Blue™ IL-1R cells endogenously express the human IL-1 receptor and were stably transfected with the murine IL-1 receptor rendering these cells very sensitive to both human and murine IL-1α and IL-1β. HEK-Blue™ IL-1R cells express a SEAP reporter gene under the control of the IFN-β minimal promoter fused to five NF-κB and five AP-1 binding sites. Binding of IL-1β to its receptor IL-1R on the surface of HEK-Blue™ IL-1R cells triggers a signaling cascade leading to the activation NF-κB and the subsequent production of SEAP. Of note, HEK-Blue™ IL-1R derive from HEK-Blue™ IL-1β cells in which the TNF-α response is blocked while the response to murine TNF-α remains intact.
Detection of SEAP in the supernatant of HEK-Blue™ IL-1R cells can be readily assessed using QUANTI-Blue™ Solution, a SEAP detection medium. QUANTI-Blue™ Solution turns blue in the presence of SEAP which can be easily quantified using a spectrophotometer.
FEATURES OF HEK-BLUE™ IL-1R CELLS:
- Fully functional IL-1 signaling pathway
- Do not respond to human TNF-α
- Readily assessable SEAP reporter activity
- Functionally tested and guaranteed mycoplasma-free
APPLICATIONS OF HEK-BLUE™ IL-1R CELLS:
- Detection of human and murine IL-1α and IL-1β
- Detection of IL-1 in various biological samples including serum
- Screening of anti-IL-1α or anti-IL-1β antibodies
1. Dinarello C., 2018. Overview of the IL-1 family in innate inflammation and acquired immunity. Immunol Rev. 281(1): 8–27.
2. Lopez-Castejon G. & Brough D., 2011. Understanding the mechanism of IL-1β secretion. Cytokine Growth Factor Rev. 22(4):189-95.
3. Wiggins K.A. et al., 2019. IL-1α cleavage by inflammatory caspases of the noncanonical inflammasome controls the senescence-associated secretory phenotype. Aging Cell. 18(3):e12946.
4. Weber A. et al., 2019. Interleukin-1 (IL-1) pathway. Sci Signal. 3(105):cm1.
Response of HEK‑Blue™ IL‑1R cells to a panel of cytokines. Cells were stimulated with various human and murine recombinant cytokines: 1 ng/ml of hIL-1β, mIL-1β, hIL-1α, mIL-1α, 10 ng/ml of hIL-18, hIFN-γ, hTNF-α, mTNF-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.
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™
Detection range for human IL-1β: 0.1 pg - 100 ng/ml
Detection range for human IL-1α, murine IL-1α, and murine IL-1β: 1 pg - 100 ng/ml
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- 1 vial containing 3-5 x 106 cells
- 1 ml of Hygromycin B Gold (100 mg/ml)
- 1 ml of Puromycin (10 mg/ml)
- 1 ml of Zeocin™ (100 mg/ml)
- 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)
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