The relative permeabilities for sodium, potassium and chloride in guinea pig mammary gland slices are determined by means of ion flux studies with radioisotopes. 2. Assuming that there are no significant electrogenic potential components, we calculate permeability ratios PNa/PK = 0.97 and Pc1/PK = 1.25. 3. Substitution of these values in the Goldman equation yields membrane potentials of--15 mV before and--13 mV after ouabain treatment. 4. This small change in membrane potential explains the absence of a significant change in chloride content upon ouabain application, which leads to large changes in intracellular sodium and potassium concentrations.