Wire myography is a widely used method to study the influence of vasoactive agents on the vascular tone and the contractile properties of blood vessels ex vivo. The literature has several papers with regard to the theoretical background and basic guidelines of myography. Here, we propose a highly detailed practical guide for studying the functional responses of mouse aorta to pharmacological stimuli that modify vascular tone. We have included photographs taken during the dissection and mounting of aortic rings, screenshots from representative LabChart diagrams, as well as a table with “classical” reagents used to probe vascular reactivity, with information on their mechanism of action and the range of concentrations commonly used.