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用ＮＰ－４０处理牛精子获得膜抽提液，经伴刀豆素Ａ－Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析，分离出与伴刀豆素Ａ结合的糖蛋白，经ＳＤＳ－聚丙烯酰胺凝胶电泳显示出分子量为７２、５７和３ｌｋＤ的３条蛋白质带。将分离到的糖蛋白电转移在硝酸纤维素膜上，用酶标的伴刀豆素Ａ检测，３条蛋白质带均呈阳性。用分离的糖蛋白免疫兔，制备抗血清。用抗血清对牛精子膜ＮＰ－４０抽提物进行免疫印迹法分析，检测出３条阳性带，与亲和层析法分离的糖蛋白带相对应。用抗血清对牛精子进行免疫细胞化学标记，表明该糖蛋白主要分布于精子顶体区。
The bovine spermatozoa were treated with NP-40 to obtain the membrane extract, and the glycoprotein bound to concanavalin A was isolated by affinity chromatography with concanavalin A-Sepharose 4B. SDS-polyacrylamide gel electrophoresis Three protein bands with molecular weights of 72, 57 and 3lkD. The isolated glycoproteins were electrotransferred on nitrocellulose membrane and detected with concanavalin A, and all three protein bands were positive. Rabbits were immunized with isolated glycoproteins to prepare antisera. Immunoblot analysis of NP-40 extracts from bovine sperm membranes with antiserum detected three positive bands corresponding to glycoprotein bands separated by affinity chromatography. Immunocytochemical labeling of bovine sperm with antiserum showed that the glycoproteins were mainly distributed in the sperm acrosome.