A previous study reported that the uptake and release kinetics of ouabain by human erythrocytes in suspension could well be explained by a physical model which involves the slow Langmuir binding of the drug to the erythrocyte membrane. The purpose of the present investigation was to assess quantitatively the thermodynamics of this drug-membrane receptor interaction in order to evaluate the consistency of these parameters with the proposed kinetics model. Cellular drug uptake and release experiments were conducted at 20, 30 and 40°C, and the Langmuir adsorption and desorption rate constants as well as the Langmuir adsorption isotherms determined from the rate data. With the knowledge of these Langmuir parameters, it was possible to estimate the magnitude of all relevant thermodynamic properties by the use of established physicochemical theories. The activation energies and entropies for the ouabain adsorption and desorption processes were computed as 105 kJ/mol, 231 J/K per mol, 180 kJ/mol and 245 J/K per mol, respectively. The kinetic and isosteric heats of adsorption were found to be −75.0 and −72.4 kJ/mol, respectively. These findings suggest that the ouabain-erythrocyte membrane interaction represents a case of activated chemisorption which follows the Langmuir isotherm, thus, further underscoring the appropriateness of the Langmuir binding kinetics model.