Our knowledge of nitric oxide (NO) as a crucial endogenous signalling molecule continues to expand. Many, but not all, of the actions of NO are mediated by activation of soluble guanylyl cyclase (sGC) in target tissues. The aim of this chapter is to encapsulate the functions of NO in mammalian biology, tied to the chemistry of this unusual signalling entity. The experimental usefulness and therapeutic potential of the most widely utilised NO donor drugs is reviewed, with special consideration given to the importance of choosing the correct NO donor for any given experiment, in vitro, in vivo or in clinical studies.