【摘要】 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。更多还原

【Abstract】 Plasma membrane glycoproteins of bovine sperm were solubilized with Nonidet P-40 and isolated by affinity chromatography on concanavalin A-Sepharose 4B column. The isolated concanavalin A-binding glycoprotein was subjected to SDS-PAGE and exhibited three bands with molecular weights of 72, 57 and 3l kD, After trasfering the obtained glycoproteins on nitrocellulose membrane by Westernblotting ,all the three bands were labeled by HRP-concanavalin A.Antiserum against the glycoprotein was raised in rabbits. Using immunoblotting technique with the antiserum, three positive bands were detected in the Nonidet P-40 extract of sperm membrane. These bands were in corespondence with the results of SDS-PAGE of the glycoprotein.Using immunocytochemical method with the antiserum,strong reactivity was observed on the acrosomal region of the bovine sperms. When sperms were stained by HRP-labeled concanavalin A following antiserum treatment,only weak staining was found on the acrosomal region. The result indicates that concanavalin A-binding glycoproteins mainly locate on the acrosomal region.更多还原