
Abstract Covalent fluorescent probes have emerged as versatile chemical tools to visualise active enzymes in complex biological systems. When tailored for specific applications, ranging from activity‐based protein profiling for drug development to high spatiotemporal resolution imaging of enzymatic activities, these probes provide unique insights into rapid lipid metabolism. The recent development of reverse‐designing optimised inhibitors into probes, together with improved analytical techniques, is rapidly advancing our ability to resolve enzyme function in great detail. In this mini‐review, we summarise the current landscape of covalent fluorescent probes for the 2‐arachidonoylglycerol (2‐AG) metabolising enzymes (diacylglycerol lipase, monoacylglycerol lipase, ABHD6 and ABHD12), highlighting the discoveries they have enabled and discussing strategies to address gaps where probes remain lacking.
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