分次灌胶制备二维凝胶电泳大胶  

Preparation of large gel of 2-D polyacrylamide gel electrophoresis with multiple times’gel potting

作　　者：徐胡斌 朱茵 刘安敏 赵起超 谢恬[1,2] 陈功星 XU Hubin;ZHU Yin;LIU Anmin;ZHAO Qichao;XIE Tian;CHEN Gongxing(School of Medicine,Hangzhou Normal University,Hangzhou 311121,China;Engineering Lab of Development and Application of Traditional Chinese Medicine of Zhejiang Province,Hangzhou 311121,China)

机构地区：[1]杭州师范大学医学部,浙江杭州311121 [2]浙产中药材资源开发与应用浙江省工程实验室,浙江杭州311121

出　　处：《华夏医学》2021年第2期18-21,共4页Acta Medicinae Sinica

基　　金：国家自然科学基金重点项目(81730108);浙江省科技厅公益性技术应用研究项目(LGF18H290002)。

摘　　要：目的:探索解决二维凝胶电泳大胶制备过程中的漏胶问题。方法:分离人肺腺癌细胞株A549蛋白,采用3次与单次灌胶制备的大胶板进行二维电泳,然后,考马斯亮蓝染色与扫描分析。结果:3次灌胶制备大胶成功率90%,高于单次灌胶制胶成功率20%;采用分次灌胶的大胶进行二维凝胶电泳,分离的蛋白点清晰、形态良好。结论:分次灌胶能防止聚丙烯酰胺凝胶制备大胶时漏胶,制作的大胶可用于二维凝胶电泳分析。Objective:To investigate a simple and convenient method of leakage proof for preparation of large gel of 2-D polyacrylamide gel electrophoresis.Methods:The protein samples of human lung adenocarcinoma cell line A549 were abstracted by two-dimensional electrophoresis on the large gel plates prepared by 3 times and one time’s gel potting.Then,the protein samples were stained by Coomassie Blueand scanned.Results:The success rate for 3 times’gel potting was 90%,which was higher than 20%for the one time’s gel potting.When the preparation of large gel of 2-D polyacrylamide gel electrophoresis was carried out by multiple times’gel potting,the isolated protein spots were clear and were in good shape.Conclusion:The way of multiple times’gel potting can prevent the polyacrylamide gel from leaking during the preparation of large gel,and the large gel can be used for 2-D gel electrophoresis analysis in proteomics.

关 键 词：二维凝胶电泳 漏胶 电泳大胶 蛋白组学 

分 类 号：R446.1[医药卫生—诊断学]