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

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.