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2 protocols using anti human cleaved il 1β a5208206

1

NLRP3 Inflammasome Activation Pathway

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MSU, nigericin, ATP, PMA, BHB, DL-sulforaphane, poly(dA:dT), and glucose were purchased from Sigma-Aldrich. MCC950 was acquired from Selleck. BAY 11-7082, isoliquiritigenin, parthenolide, flufenamic acid, and mefenamic acid were acquired from Topscience. Human recombinant insulin was bought from Novo Nordisk. The One Touch Ultra Blood glucose Test System kit was bought from Roche. Ultrapure LPS, Pam3CSK4, MitoTracker, and MitoSOX were obtained from Invitrogen. Protein G agarose and streptavidin-coated beads were respectively supplied by Millipore and Pierce Biochemicals. ATP-coupled beads were from BioWorld Company. Anti-mouse caspase-1 (p20; AG-20B-0042) and anti-NLRP3 (AG-20B-0014) were from Adipogen. Anti-ASC (sc-22514-R) and anti-NEK7 (SC-50756) were from Santa Cruz. Anti-β-actin (P30002) was bought from Abmart. Anti-human caspase-1 was from Cell Signaling Technology. Anti-human cleaved IL-1β (A5208206) was from Sangon Biotech. Anti-mouse IL-1β (p17; AF-401-NA) was from R&D Systems. Anti-Flag (F2555) and anti-Mcherry (V4888) were from Sigma-Aldrich. Recombinant human NLRP3 was from Novus Biologicals. A standard HFD (D12492, 60% kcal fat) was from Research Diet Company.
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2

NLRP3 Inflammasome Activation and IL-1β Regulation

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ATP, MSU, poly A:T, nigericin, and glucose were from Sigma. The human recombinant insulin was from Nova Nordisk. One Touch® Ultra® Blood glucose Test System was from Roche. Oridonin (s2335) was bought from Selleck. MitoSOX, ultrapure LPS, Pam3CSK4 and MitoTracker were from Invitrogen. Protein G agarose and streptavidin-coated beads were, respectively, supplied by Millipore and Pierce Biochemicals. Anti-Flag (F2555) and anti-VSV (V4888) were bought from Sigma. Anti-NLRP3 (AG-20B-0014) and anti-mouse caspase-1 (p20) (AG-20B-0042) antibodies were from Adipogen. Anti-NEK7 (SC-50756) and anti-ASC (sc-22514-R) antibodies were obtained from Santa Cruz. Anti-β-actin (P30002) was bought from Abmart. Anti-human caspase-1 was from Cell Signaling Technology. Anti-human cleaved IL-1β (A5208206) was from Sangon Biotech. Anti-mouse IL-1β (p17) (AF-401-NA) was from R&D Systems. A standard high-fat diet (D12492, 60% kcal fat) was from Research Diet Company.
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