Non-specific ligand accumulation into membrane material, which may contribute considerably to the experimental signal obtained in binding studies with labelled amphipathic and hydrophobic ligands, may be accounted for by linear partition of the ligands into the membrane phase. For application to binding data obtained at a single membrane-lipid concentration, a fitting procedure is proposed which allows one to correct for non-specific ligand partition. If the assumption is met that the amount of acceptor-bound ligand is small compared to the total amount present in the system, one can validly interpret the data in terms of total ligand concentrations. The apparent dissociation constants Kd(app) thus obtained should be corrected for the often large effects of the size of the partition compartment(s), by performing assays at several membrane-lipid concentrations. The importance of the latter correction is stressed and an approach for obtaining the characteristic effective dissociation constants Kd' is indicated. The procedure also yields estimates of the ligand/membrane partition coefficients.