Calcium-binding proteins (CaBPs) have been described as involved in the stimulus-secretion coupling mechanisms in secretory glands. CaBPs were revealed with 45Ca, after electrophoresis in SDS-PAGE and transference to Zeta probe membranes, in Duvernoy's or venom gland homogenates from three families of South American snakes: Viperidae (Bothrops jararaca and Crotalus durissus terrificus); Elapidae (Micrurus corallinus), and Colubridae (Phylodrias patagoniensis and Oxyrhopus trigeminus). A band with an estimated molecular weight of 12 KDa was found in all glands studied. Bands with 17, 28, and 67 KDa were found in all glands, except in O. trigeminus Duvernoy's gland. A 18 KDa band was found in Viperidae and Elapidae venom glands, and a 88 KDa band was observed only in Viperidae venom gland homogenates. Some of these CaBPs were identified by Western blotting or by immunohistochemistry, as parvalbumin (12 KDa) and calbindin (28 KDa). When the secretion of these glands were analyzed, CaBPs were detected only in B. jararaca venom, with bands of 14, 35, 42, and 72 KDa. The profile of CaBPs was not modified at different phases of the secretory cycle of the glands, as well as after isoproterenol treatment.