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Monosodium urate (crystals)

ALX-400-047

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NLRP3 inflammasome activator

NLRP3/NALP3 inflammasome activator. Stimulates the caspase-1 activating NLRP3/NALP3 inflammasome, resulting in the production of active IL-1β and IL-18.

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Product Details

Alternative Name	MSU (crystals), Uric Acid (crystals)
Appearance	White to off white solid.
CAS	1198-77-2
Couple Target	Inflammasome
Couple Type	Activator
Formula	C₅H₃N₄O₃ . Na
Identity	Determined by IR.
MW	190.1
RTECS	YU7062000
Solubility	Soluble in PBS or 1M NaOH (25 mg/ml). Solution may appear as a slightly hazy suspension.

Handling & Storage

Use/Stability	As indicated on product label or CoA when stored as recommended.
Handling	Vortex before use.
Long Term Storage	-20°C
Shipping	Ambient Temperature

Regulatory Status	RUO – Research Use Only

Oxidized DNA fragments exit mitochondria via mPTP- and VDAC-dependent channels to activate NLRP3 inflammasome and interferon signaling: Xian, H., Watari, K., et al.; Immunity 55, 1370 (2022), Abstract
Statins repress needle-like carbon nanotube- or cholesterol crystal-stimulated IL-1β production by inhibiting the uptake of crystals by macrophages: H. Cui, et al.; Biochem. Pharmacol. 188, 114580 (2021), Abstract
Metformin inhibition of mitochondrial ATP and DNA synthesis abrogates NLRP3 inflammasome activation and pulmonary inflammation: Xian, H., Liu, Y., et al.; Immunity 54, 1463 (2021), Abstract
Sensing low intracellular potassium by NLRP3 results in a stable open structure that promotes inflammasome activation: Tapia-Abellán, A., Angosto-Bazarra, D., et al.; Sci. Adv. 7, eabf4468 (2021), Abstract
Early endosome autoantigen 1 regulates IL-1β release upon caspase-1 activation independently of gasdermin D membrane permeabilization: A. Baroja-Mazo, et al.; Sci. Rep. 9, 5788 (2019), Abstract — Full Text
Choline Uptake and Metabolism Modulate Macrophage IL-1β and IL-18 Production: Sánchez-López, E., Zhong, Z., et al.; Cell Metab. 29, 1350 (2019), Abstract
P2Y6 Receptor Signaling Pathway Mediates Inflammatory Responses Induced by Monosodium Urate Crystals: H. Uratsuji, et al.; J. Immunol. 188, 436 (2012), Abstract
Monosodium urate crystals in inflammation and immunity: Y. Shi, et al.; Immunol. Rev. 233, 203 (2010), Abstract
Inflammasome activation in NADPH oxidase defective mononuclear phagocytes from patients with chronic granulomatous disease: F. Meissner, et al.; Blood 116, 1570 (2010), Abstract — Full Text
Uric Acid-Driven Th17 Differentiation Requires Inflammasome-Derived IL-1 and IL-18: C. Conforti-Andreoni, et al.; J. Immunol. 187, 5842 (2010), Abstract

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Datasheet, Manuals, SDS & CofA

Manuals And Inserts

Datasheet

Specific Protocol

Example:

Activation of THP-1 cells by uric acid:

THP-1 cells are cultured in RPMI 1640 supplemented with 10% heat inactivated fetal bovine serum (FCS), 50μM β-mercaptoethanol, 2mM L-glutamine and penicillin/streptomycin. THP-1 cells are grown in suspension to a density of 0.7-1.5x10⁶ cells/ml in tissue culture flasks.
Treat THP-1 cells for 3 hours with 25ng/ml PMA (Prod. No. BML-PE160) the day before stimulation.
Wash cells with PBS and plate them in a 12-well plate at a density of 6x10⁵ cells/well (or 6-well plate at density of 1x10⁶ cells/well) in RPMI medium (this increases the phagocytic properties of the cells and induces a constitutive production of proIL-1β).
The next day, wash cells with PBS and add 500µl Optimem (Invitrogen) in 12-well plate or 1mL in 6-well plate.
Stimulate the cells for 6 hours by addition of 50-200µg/ml of the suspension of uric acid (vortex the suspension just before use).
As a positive control, 50µg/ml of LPS from E. coli, Serotype R515 (Re) (TLRgrade™) (liquid) (Prod. No. ALX-581-007) are incubated for 6 hours. (50µg/ml LPS showed very strong stimulating response, 10µg/ml could be used for future test)

Detection of the mature IL-1β in the supernatant of uric acid-activated THP-1 cells:

Collect the supernatant of each well, vortex and centrifuge for 5 min. at 13'000 rpm, then save the supernatant.
Detection of IL-1β in the supernatant of uric acid-activated THP-1 cells by ELISA (Prod. No. ADI-900-130).

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