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SiO2 nanoparticles; NLRP3 inflammasome inducer
20 mg (2 x 10 mg)
NLRP3 Inflammasome Inducer
SiO2 nanoparticles (Nano-SiO2) are single particles of silica dioxide, an inorganic metal oxide, with a diameter of less than 100 nm. Several studies have demonstrated that Nano-SiO2 triggers interleukin-1β (IL-1β) secretion in vitro and in vivo [1, 2]. IL-1β is produced as a pro-protein which is proteolytically processed to its active form by caspase-1. The secretion of IL-1β is an indicator of the NLRP3 inflammasome induction.
The NLRP3 inflammasome is an intracellular multi-protein complex that plays a central role in innate immunity [3, 4]. It is activated by a two-step process; a first signal (‘priming’) is provided by microbial molecules such as lipopolysaccharide (LPS), while the second signal is provided by a wide array of stimuli including bacterial toxins, endogenous molecules, crystals or nanoparticles such as Nano-SiO2. This triggers inflammasome multimerization and caspase-1 activation with the subsequent maturation and secretion of IL-1β and IL-18. Research has confirmed that the IL-1β secretion and pro-inflammatory activity of Nano-SiO2 are mediated by the NLRP3 inflammasome [1, 2].
InvivoGen’s Nano-SiO2 is designed for in vitro assays. Its ability to induce the NLRP3 inflammasome has been validated using THP-1 Null cells.
Features of Nano-SiO2:
- Potent inducer of the NLRP3 inflammasome
- Nanoparticles with a diameter of <100 nm
- Each lot is functionally tested
1. Nakayama M. et al., 2018. Macrophage recognition of crystals and nanoparticles. Front Immunol. 9:103.
2. He Y. et al., 2016. NEK7 is an essential mediator of NLRP3 activation downstream of potassium efflux. Nature. 530(7590):354-7.
3. Li H. et al., 2008. Cutting Edge: Inflammasome activation by Alum and Alum’s adjuvant effect are mediated by NLRP3. J Immunol. 181:17-21.
4. Hornung V. et al., 2008. Silica crystals and aluminium salts activate the NALP3 inflammasome through phagosomal destabilization. Nature Immunol. 9:847-856.
IL-1β production in THP1-Null cells.
THP1-Null cells, primed with LPS (1 µg/ml for 3h), were stimulated with increasing concentrations of Nano-SiO2. After overnight incubation, IL-1β secretion was analyzed by adding 50 µl of supernatant from treated THP1-Null cells to HEK-Blue™ IL-1β cells. IL-1β-induced activation of NF-κB was assessed by measuring the levels of SEAP in the supernatant of HEK-Blue™ IL-1β cells using QUANTI-Blue™ Solution, a SEAP detection reagent, and by reading the optical density (OD) at 630 nm.
Working concentration: 10-250 µg/ml
CAS number: 7631-86-9
Linear formula: SiO2
Molecular weight: 60.08 g/mol
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- 20 mg (2 x 10 mg) Nano-SiO2 (nanoparticles of silica dioxide)
Nano-SiO2 is shipped at room temperature.
Store at room temperature.Back to the top
THP-1/HEK-Blue™ IL-1β Assay
Nano-SiO2 can be used to induce the NLRP3 inflammasome in cellular assays, such as InvivoGen’s THP1/HEK-Blue™ IL-1β assay. This assay uses the secretion of IL-1β by THP1-Null cells as an indicator of NLRP3 inflammasome induction. The production IL-1β by these cells is measured using HEK-Blue™ IL-1β cells.
1- Production of IL-1β by THP-1 cells: Typically, THP1-Null cells are primed with lipopolysaccharide (LPS). This treatment induces the production of NLRP3 and pro-IL-1β, the immature form of IL-1β. Subsequent stimulation with inflammasome inducers, such as Nano-SiO2, 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: THP1-Null cell supernatants containing IL-1β can be added to HEK-Blue™ IL-1β cells. This activates NF-κB and leads to the subsequent production of secreted embryonic alkaline phosphatase (SEAP). The presence of SEAP in HEK-Blue™ IL-1β supernatants is assessed using QUANTI-Blue™ Solution, a SEAP detection medium.Back to the top