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本文通过标记完整精子的表面膜蛋白,再使用等电聚焦(IEF)和非平衡pH梯度胶体电泳(NEPHGE)研究精子蛋白提取物。 正常人精液,Percoll纯化为活动精子数超过90％的精子悬液。加Iodo珠和~(125)I-Na,20℃振摇培养10min,进行表面蛋白的放射碘标记;或加NHS-LC生物素37℃振摇共育10min,进行表面蛋白的生物素标记。标记后,用Percoll作再纯化,洗涤,所获精子仍为顶体完整精子。由碘标记精子的放射自显影,
In this paper, sperm protein extracts were studied by labeling the surface membrane proteins of intact sperm, followed by isoelectric focusing (IEF) and non-equilibrium pH gradient gel electrophoresis (NEPHGE). Normal human semen, Percoll purified to more than 90% of sperm motility sperm suspension. Iodo beads and ~ (125) I-Na were added and shake-cultured at 20 ° C for 10 min for iodine labeling of surface proteins. The biotinylated protein was labeled with NHS-LC biotin for 10 min at 37 ° C. After labeling, the cells were repurified and washed with Percoll, and the spermatozoa obtained were still intact acrosome spermatozoa. Autoradiography by iodine-labeled sperm,