Na/K ATPase activity in rat myometrial cells in culture exhibited a Kapp of 0.93 mM for Rb+ and a Ki of 31 microM for ouabain with respect to Rb+. 2. 86Rb+ uptake was stimulated by serum and monensin but was not affected by the uterine relaxants isoproterenol and relaxin in 0.5-7.5 mM Rb+. Nonetheless, these relaxants elicited significant increases in 45Ca2+ efflux under similar conditions. 3. These data suggest that increased Na/Ca exchange resulting from a stimulation of Na/K ATPase is not involved in the mechanism of action of relaxin and isoproterenol in the uterus.