Antibodies raised against rabbit sperm membrane protein (rSMP-B) interacted with a rat testis cytosolic protein with an estimated molecular weight of 70 kD. The clone expressing rSMP-B was isolated by epitope selection from a rat testis lambda gt11 vector cDNA library. An Eco R1-cut 2.0 kb cDNA fragment encoding rSMP-B was identified. The cDNA fragment (RSD-1) was isolated, subcloned and restriction mapped. Dot hybridization analysis of mRNA prepared from human, rabbit and rat testes showed that the gene encoding rSMP-B is expressed in the testes of all three mammals.