Mammalian spermatozoa fertilize only after capacitation. The removal of decapacitation factors that inhibit the acrosome reaction (AR) is one of the events taking place during capacitation. In this report, human sperm were capacitated by 18-h incubation in Biggers, Whitten & Whittingham medium (BWW) medium and the proteins, on release, were analysed. After gel filtration by high-performance liquid chromatography a main peak with an approximate native molecular weight of 130 kDa was recognized by an antinormal seminal plasma antibody. This fraction was able to inhibit the follicular fluid as well as the progesterone-induced AR, when added to capacitated spermatozoa. Additionally, it reacted with an antibody directed against seminal plasma from vasectomized donors but not with an antibody against epididymal proteins. The AR inhibitory activity was heat-denatured, could be partially destroyed when treated with proteases, and bound to Concanavalin-A and wheat germ lectins. These results suggest that during in vitro capacitation, human spermatozoa release a glycoproteic decapacitation factor produced by accessory sex glands.