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measure non-precipitating antibodies. The more recently developed cytotoxic tests (Hamerlynck & R\l=u"\mke,1968) also detect a limited spectrum of antibody types. Binding tests which measure antibody absorbed to a target cell have rarely been used in studies of sperm antigens. Noyes (1969) used a direct binding test for human anti-sperm antibodies which involved labelling a patient's sera with 131I. In the work described here, an isotopic antiglobulin technique (Harder & McKhann, 1968; Bomford, Breitner, Mitchison, Negroni & Raff, 1969; Sparks, Ting, Hammond & Herberman, 1969) has been adapted for use with mouse spermatozoa. This is an indirect or `sandwich' technique which utilizes 125I-rabbit-anti-mouse-\g=g\-globulinto detect mouse anti-mouse-spermatozoa antibodies bound to spermatozoa. The one radioactive reagent can be used with many different mouse antisera and, thus, obviates the need to iodinate each separately, a requirement of the direct technique. The mouse anti-mouse-spermatozoa antisera used in the development of the test were hyperimmune antisera prepared in random-bred Qstrain female mice using an 8-week course of thrice weekly injections of spermatozoa (Bell & McLaren, 1970). Serum from mice immunized with whole spermatozoa (Group Sp) or the supernatant fraction of disrupted spermatozoa (Group Sup+ Sup —A) were pooled and designated Pool S and T-34, respectively. A rabbit anti-mouse-y-globulin was prepared by immunization of rabbits with the purified y-globulin from non-immune CBA mice prepared by ammonium sulphate fractionation and DEAE cellulose ion-exchange chromatography (Fahey, 1967). The resultant rabbit anti-mouse-y-globulin serum was carefully purified before labelling by an immunoadsorbent column. The easily purified

AN ANTIGLOBULIN TEST FOR THE DETECTION OF SURFACE ANTIGENS OF MOUSE SPERMATOZOA