Muscarinic receptor and beta-adrenergic receptor binding were measured simultaneously in a membrane fraction of bovine tracheal smooth muscle using [3H]-L-quinuclidinyl benzilate and [125I]-(-) iodocyanopindolol. The binding characteristics, affinity and receptor density, obtained in the double receptor assay and in the control experiments were the same within experimental error. Moreover, there appears to be neither a significant influence of an excess of d,1-propranolol on [3H]-L-quinuclidinyl benzilate binding nor a significant influence of an excess of 1-quinuclidinyl benzilate on [125I]-(-)iodocyanopindolol binding. The method is advantageous where both receptors have to be assayed and where limited amounts of biological material, like in biopsy specimen, are available.