A practical guide to working with H₂S at the interface of chemistry and biology

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Hydrogen sulfide (H₂S) is the most recently accepted endogenously produced gasotransmitter and is now implicated in a variety of physiological functions. In this tutorial review, our goal is to provide researchers new to the field of H₂S chemical biology with practical considerations, pitfalls, and best practices to enable smooth entry into investigations focused on biological H₂S. We present practical handling and safety considerations for working with this reactive biomolecule, and cover basic roles of H₂S biogenesis and action. Experimental methods for modulating H₂S levels, including enzymatic knockout, RNA silencing, enzymatic inhibition, and use of small molecule H₂S donors are highlighted. Complementing H₂S modulation techniques, we also highlight current strategies for H₂S detection and quantification.