人精子膜抗原的提取及分析

目的:用两种酶裂解法提取精子膜抗原并对其进行蛋白浓度测定和成分分析。方法:分别用尿素裂解液和表面活性剂Triton X-100﹑脱氧胆酸钠(DOC)的TDOC裂解液裂解液提取精子膜抗原,借助BCA蛋白浓度测定试剂盒测定蛋白浓度,并通过SDS-PAGE法比较分析蛋白成分。结果:两种方法提取的精子膜抗原经SDS-PAGE,考马斯亮蓝染色后均有条带显示,TDOC裂解液提取的样本中显示仅有一条蛋白带,分子质量集中在100ku左右,尿素裂解液提取的样本中有多条蛋白条带,分子质量集中在130～35ku。结论:尿素裂解液提取法提取精子膜抗原的效果优于TDOC裂解液提取法,为抗精子相关抗体的研究提供了条件。

Assessment of the approach to human sperm membrane antigen extraction

Objective：To assess the two approaches to extracting human sperm membrane antigen by enzymatic lysis through determining the pro- tein concentration and its fraction. Methods ： Extraction of the sperm anti- gen was carried out respectively with urea lysis buffer plus Surfactant Triton X-100 and sodium deoxycholate （DOC） TDOC lysls buffer. BCA protein assay kit was used to determine the protein concentration, which was further verified with SDS-PAGE method. Results： Although the two techniques exposed strips for sperm membrane antigen after SDS-PAGE treatment and Coomassie brilliant blue staining, yet only one band was seen at 100 ku for the sample treated with TDOC lysis buffer, whereas more strips were observed at 130 ku to 35 ku for the one managed with u- rea lysis buffer. Conclusion ： Extraction of the sperm membrane antigen by urea lysis buffer seems better than by TDOC lysis buffer,and this findings will have certain implications in further study of anti-sperm antibodies.