目的　精子蛋白双向电泳的建立 ,并利用其初步研究冻融前后精子蛋白的改变 ,为进一步筛选差异表达的蛋白奠定基础。方法　来源于同一个人的冻融前后精子样品经溶解液溶解 ,并以等电聚焦 (IEF)作为第一向电泳 ,以变性的聚丙烯酰胺凝胶电泳 (SDS -PAGE)作为第二向 ,对上述样品进行分离 ,凝胶银染后进行比较。结果　建立了精子蛋白的双向电泳 ,并用其分析了冻融前后精子蛋白的改变。结论　冻融前后精子蛋白发生不止一种蛋白的改变 ,本研究的实验方法是一种研究精子冻融蛋白改变的有效手段 ,但仍有待进一步优化 Objective To establish the two-dimensional gel electrophoresis of sperm protein and to make preliminary studies on the changes of sperm proteins before and after freezing-thawing so as to lay the foundation for the further screening of differentially expressed proteins. Methods Before and after freezing and thawing, the sperm samples from the same individual were dissolved by dissolving solution. The isoelectric focusing (IEF) was used as the first electrophoresis and the denatured polyacrylamide gel electrophoresis (SDS-PAGE) as the second direction. The above samples were separated and compared by gel silver staining. Results The two-dimensional electrophoresis of sperm protein was established and used to analyze the changes of sperm protein before and after freezing-thawing. Conclusion There is more than one protein change in sperm protein before and after freezing and thawing. The experimental method in this study is an effective method to study the change of sperm freezing and thawing protein, but it needs further optimization