Monosodium Urate CAS 1198-77-2

CAS Number

1198-77-2

Grade

Highly Purified

Molecular Formula

C5H3N4O3Na

Molecular Weight

190.09

EU Commodity Code

38220090

Shipping Temp

Blue Ice

Storage Temp

-20°C

7,9-Dihydro-1H-Purine-2,6,8(3H)-trione; 1H-Purine-2,6,8-triol; 2,6,8-Trihydroxypurine; 2,6,8-Trioxopurine; 2,6,8-Trioxypurine; Lithic Acid; NSC 3975; Purine-2,6,8(1H,3H,9H)-trione

NLRP3/NALP3 inflammasome activator. Specially crystallized and tested for biological activity. Stimulates the caspase-1 activating NLRP3/NALP3 inflammasome, resulting in the production of active IL-1beta and IL-18.

Synonyms

7,9-Dihydro-1H-Purine-2,6,8(3H)-trione; 1H-Purine-2,6,8-triol; 2,6,8-Trihydroxypurine; 2,6,8-Trioxopurine; 2,6,8-Trioxypurine; Lithic Acid; NSC 3975; Purine-2,6,8(1H,3H,9H)-trione

CAS No

1198-77-2

Molecular Formula

C5H3N4O3Na

Molecular Weight

190.09

Appearance

Supplied as a liquid in PBS. Ready-to-use solution and is completely sterile and endotoxin-free.

Purity

≥95%

Sterility

Sterile

Endotoxin

None detected

Solubility

PBS

Storage and Stability

May be stored at RT for short-term only. Long-term storage is recommended at -20°C. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Activation of THP-1 cells by uric acid

1. THP-1 cells are cultured in RPMI 1640 supplemented with 10% heat inactivated fetal bovine serum (FCS), 50uM β-mercaptoethanol, 2mM L-glutamine and penicillin/streptomycin. THP-1 cells are grown in suspension to a density of 1.5x10e6 cells/ml in tissue culture flasks. 2. Treat THP-1 cells for 3 hours with 0.5uM PMA the day before stimulation. 3. Wash cells with PBS and plate them in a 12-well plate at a density of 6x10e5 cells/well in RPMI medium. 4. The next day, wash cells with PBS and add 500ul Optimem (Invitrogen). 5. Stimulate the cells for 6 hours by addition of 50-300ug/ml of the suspension of uric acid (vortex the suspension just before use). 6. As a positive control, 5mM of ATP is incubated for 6 hours.

Detection of the mature Caspase-1 in the supernatant of uric acid-activated THP-1 cells

1. Collect the supernatant of each well (500ul). Add 500ul methanol and 125ul chloroform. 2. Vortex and centrifuge for 5 min at 13,000 rpm. 3. Remove the upper phase. 4. Add 500ul methanol and centrifuge for 5 min at 13,000 rpm. 5. Remove the supernatant, dry the pellet for 5 min at 50°C. 6. Add 60ul of 1X loading buffer and DTT (final concentration of 0.1M). 7. Mix and boil before loading on a 15% SDS-gel and transfer onto nitrocellulose membrane. The mature Caspase-1 is detected using the Caspase-1 (p20) (human), mAb (Bally-1) antibody (Cat No 044627).

Storage and Stability

Short-term Storage: +4°C Long-term Storage: -20°C Stable for at least 1 year after receipt when stored at -20°C.

References

1. Monosodium urate crystals in inflammation and immunity: Y. Shi, et al.; Immunol. Rev. 233, 203 (2010)|2. Crystals of monosodium urate monohydrate enhance lipopolysaccharide-induced release of interleukin 1 beta by mononuclear cells through a caspase 1-mediated process: E.J. Giamarellos-Bourboulis, et al.; Ann. Rheum. Dis. 68, 273 (2009)|3. Malarial hemozoin activates the NLRP3 inflammasome through Lyn and Syk kinases: M.T. Shio, et al.; PLoS Pathog. 5, e1000559 (2009)|4. Gout-associated uric acid crystals activate the NALP3 inflammasome: F. Martinon, et al.; Nature 440, 237 (2006)

USBio References

No references available

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045039 Monosodium Urate (MSU), ready-to-use liquid CAS: 1198-77-2

CAS Number

Grade

Molecular Formula

Molecular Weight

EU Commodity Code

Shipping Temp

Storage Temp

7,9-Dihydro-1H-Purine-2,6,8(3H)-trione; 1H-Purine-2,6,8-triol; 2,6,8-Trihydroxypurine; 2,6,8-Trioxopurine; 2,6,8-Trioxypurine; Lithic Acid; NSC 3975; Purine-2,6,8(1H,3H,9H)-trione

References

USBio References

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