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

Hydrogen sulfide (H 2 S) 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 H 2 S chemical biology with practical considerations, pitfalls, and best practices to enable smooth entry into investigations focused on biological H 2 S. We present practical handling and safety considerations for working with this reactive biomolecule, and cover basic roles of H 2 S biogenesis and action. Experimental methods for modulating H 2 S levels, including enzymatic knockout, RNA silencing, enzymatic inhibition, and use of small molecule H 2 S donors are highlighted. Complementing H 2 S modulation techniques, we also highlight current strategies for H 2 S detection and quantification. Current methods for modulating and measuring biological H 2 S, as well as practical considerations for working with H 2 S are provided.