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2 protocols using pge2 g d5

1

Lipid Mediator Quantification Protocol

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The primary reference materials for AEA, PEA, OEA, PE2, PGE2, PGD2, AA, 2-AG, PGE2-G, 20-HETE-EA, 15-HETE-EA, 5,6-EET-EA, 8,9- EET-EA, 11,12- EET-EA, 14,15- EET-EA and their internal standards (ISTD); AEA-d8, PEA-d4, OEA-d4, PE2-d4, PGE2-d4, PGD2-d4, AA-d8, 2-AG-d5, and PGE2-G-d5 were purchased from Cayman Chemical (Ann Arbor, MI). Acetonitrile, ammonium acetate, ammonium hydroxide, Dulbecco's phosphate buffered saline (DPBS), formic acid, ethyl acetate, isopropanol, magnesium chloride (MgCl2), methylene chloride, HPLC grade methanol, potassium chloride (KCl), Tris–HCl Buffer, pH 7.5 and water were purchased from Fisher Scientific (Hanover Park, IL). Phenylmethanesulfonylfluoride (PMSF) was obtained from Cayman Chemical. Clean Up® C18 solid phase extraction (SPE) columns were purchased from United Chemical Technologies (Bristol, PA). Medical grade nitrogen was purchased from National Welders Supply Company (Richmond, VA).
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2

Profiling Endocannabinoid Metabolism Modulators

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2-AG, anandamide (AEA), PGE2-G, PGE2, and their deuterated analogs 2-AG-d5, AEA-d8, PGE2-G-d5, and PGE2-d4 were purchased from Cayman Chemical; stock solutions were prepared in ethanol (10 mg/mL). JZL184, a selective inhibitor of monoacylglyerol lipase, and KT182, an inhibitor of ABHD6,31 (link) were purchased from Cayman Chemical. KT109, an inhibitor of DAGLβ,29 (link),30 (link) and KT195, an inhibitor of serine hydrolase ABHD6,29 (link),30 (link) were purchased from Sigma-Aldrich. Stock solutions of enzyme inhibitors were prepared in dimethyl sulfoxide (DMSO, 10 mg/mL) and diluted to their final concentration in sterile saline with Tween 80.
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