Recent work in the field of ionophore‐based ion‐selective electrodes (ISEs) has greatly improved our understanding of the thermodynamics and kinetics that describe the response and selectivity of these sensors. These efforts resulted in the lowering of detection limits from micro‐ to subpicomolar concentrations, improvements of selectivities by many orders of magnitude, and major advancements in biocompatibility and long‐term stabilities. This chapter summarizes the current state of the art for an audience that is new to the field, introducing the basic concepts of ISE theory that replaced in recent years the empirical approach of the early ISE history. It illustrates, with specific examples, the design principles of host‐guest chemistry that have been used to develop ionophores for ISEs, and describes with a minimum of equations the recently developed sophisticated concepts for the efficient use of these ionophores in ISEs. Importantly, this chapter shows not only how ionophores are used in modern potentiometry to develop new ISEs but also illustrates how ionophore‐based potentiometry can support host‐guest chemistry by providing tools to determine thermodynamic properties of ionophores, such as stoichiometries and stabilities of their complexes, using only a minimum amount of ionophore.