Androgen-binding protein (ABP) is a testicular Sertoli cell secretory protein that acts as a carrier of androgen in the male reproductive tract. ABP has been characterized from a wide range of animal species, including man, rabbit and rat. However, it has been widely accepted that mice do not produce testicular ABP. We have used immunological and molecular biological techniques to demonstrate that the ABP gene is expressed in the CD1 mouse. Steroid-binding, radioimmunoassay and immunocytochemical studies demonstrated that ABP is present in mouse testis and epididymis, but at 1/50 to 1/25 the level of rat epididymis. A 1.7 kilobase mRNA, homologous with rat ABP cDNA, was identified in mouse testis and Sertoli cells by Northern blot hybridization, but at a much lower level than in the rat. An ABP cDNA was isolated from a mouse testis cDNA library and encoded a protein (403 residues) with 89% of the amino acid residues identical to rat ABP, including a signal peptide. Our results indicate that ABP is expressed in the mouse and past failures to detect androgen-binding activity were due to the low level of ABP protein.