HEK-Blue™ IL-1β Cells
|HEK-Blue™ IL-1β cells||Unit size||Cat. code||Docs||Qty||Price|
Human & Mouse IL-1β Reporter Cells
3-7 x 10e6 cells
Human & Mouse IL-1Beta Reporter Cells
HEK-Blue™ IL-1β cells allow to detect bioactive IL-1β by monitoring the activation of the NF-κB and AP-1 pathways. They derive from HEK-Blue™ TNF-α/IL-1β cells in which the TNF-α response has been blocked. Therefore, HEK-Blue™ IL-1β cells respond specifically to IL-1. They express a NF-κB/AP-1-inducible SEAP reporter gene. Binding of IL-1β to its receptor IL-1R on the surface of HEK-Blue™ IL-1β cells triggers a signaling cascade leading to the activation NF-κB and the subsequent production of SEAP.
HEK-Blue™ IL-1β cells are useful to monitor IL-1β in inflammasome activation studies. These cells detect the presence of IL-1β in samples which triggers the production of SEAP. Levels of SEAP can be easily monitored using QUANTI-Blue™. Stimulation of HEK-Blue™ IL-1β cells with recombinant human IL-1β can be blocked by the neutralizing monoclonal anti-hIL-1β-IgG antibody.
THP1 cells pretreated with PMA and primed with LPS (1 μg/ml) were stimulated with ATP (5 mM), nigericin (1 μM), alum (200 μg/ml), MSU (200 μg/ml) or CPPD (200 μg/ml). After 24h incubation, THP-1 supernatants or recombinant IL-1β (0.1 ng/ml) were added to HEK-Blue™ IL-1β cells. IL-1β - induced NF-kB activation was assessed by measuring the levels of SEAP in the supernatant of HEK-Blue™ IL-1β cells using the QUANTI-Blue™ assay.
Stimulation of HEK-Blue™ IL-1β cells by recombinant human and murine IL-1β and human TNF-α. After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ and reading the optical density (O.D.) at 655 nm.
|Human IL-1β||0.2 +/- 0.1 ng/ml||40|
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-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.
Anti-hIL-1β-IgA was incubated with 1 ng/ml of hIL-1β for 30 minutes prior to the addition of HEK-Blue™ 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.
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™
Detects human and murine IL-1β (α)
- Detection range for human IL-1β: 100 pg - 100 ng/ml
- Detection range for murine IL-1β: 10 ng - 1 µg/ml
- 1 vial containing 3-5 x 106 cells
- 100 μl Hygromycin B Gold (100 mg/ml)
- 100 μl Zeocin™ (100 mg/ml)
- 1 ml Normocin™ (50 mg/ml)
- 1 pouch of QUANTI-Blue™
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Interleukin-1β (IL-1β) is an important mediator of the inflammatory response to infection and injury. IL-1β binds to the type 1 IL-1 receptor (IL-1R) which requires the IL-1 receptor accessory protein (IL-1RAcP) to transduce a signal. IL-1β signaling is mediated by MyD88, IRAK-1/2 and TRAF-6 leading to the activation of NF-κB and AP-1.
Binding of IL-1β to its receptor IL-1R on the surface of HEK-Blue™ IL-1β cells triggers a signaling cascade leading to the activation NF-κB and the subsequent production of SEAP.
THP-1/HEK-Blue™ IL-1β Assay
1- Production of IL-1β by THP-1 cells: Typically, THP-1 cells are pretreated with phorbol 12-myristate acetate (PMA) to become more susceptible to inflammasome activators, then are primed with lipopolysaccharide (LPS). These treatments induce the production of pro-IL-1β, the immature form of IL-1β.
Subsequent stimulation with inflammasome inducers, such as crystals or ATP, leads to NRLP3 and caspase-1 activation resulting in IL-1β maturation and secretion.
2- Detection of IL-1β by HEK-Blue™ IL-1β cells: IL-1β-containing THP-1 supernatants are added to HEK-Blue™ IL-1β cells leading to NF-κB activation and the subsequent production of SEAP. The presence of SEAP in HEK-Blue™ IL-1β supernatants is assessed using QUANTI-Blue™, a SEAP detection medium.Back to the top