The aggregation behavior of the carboxylic ionophore, lasalocid A, has been studied in egg phosphatidylcholine vesicles by monitoring the intrinsic fluorescence of lasalocid A. Self quenching of lasalocid A fluorescence in vesicles of egg phosphatidylcholine suggests aggregation of lasalocid A. When aggregated lasalocid A is treated with increasing concentrations of lipid, there is an increase in fluorescence due to gradual reduction of self quenching on lateral dilution. This confirms the presence of loosely held non-covalent aggregates of lasalocid A in the membrane. This result is relevant in elucidating the molecular mechanism of cation transport by lasalocid A across membranes.