The endocannabinoid 2-arachidonoylglycerol (2-AG) exerts its physiological action by binding to and functionally activating type-1 (CB1) and type-2 (CB2) cannabinoid receptors. It is thought to be produced through the action of sn-1 selective diacylglycerol lipase (DAGL) that catalyzes 2-AG biosynthesis from sn-2-arachidonate-containing diacylglycerols. Different methodological approaches for measuring DAGL activity in biological samples are now available. Here, a highly sensitive radiometric assay to assess DAGL activity, by using 1-oleoyl[1-14C]-2-arachidonoylglycerol as the substrate, is reported. All the steps required to perform lipid extraction, fractionation by thin-layer chromatography (TLC), and quantification of radiolabeled [14C]-oleic acid via scintillation counting are described in detail.
Keywords: 2-arachidonoylglycerol; Diacilglycerol lipase; Radiometric assay; Thin layer chromatography.
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