凝胶电泳与蛋白质印迹技术实验方法的改进和探索  

Improvement and exploration of experimental methods of gel electrophoresis and protein imprinting technology

作　　者：廖志银[1] 霍朝霞 翁登坡[1] 赵鲁杭[1] 于晓虹[1] Liao Zhiyin;Huo Zhaoxia;Weng Dengpo;Zhao Luhang;Yu Xiaohong(Basic Medicine Experimental Teaching Center,Medical College,Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学医学院基础医学实验教学中心,浙江杭州310058

出　　处：《现代盐化工》2022年第6期22-25,共4页Modern Salt and Chemical Industry

基　　金：医学院分子医学自主创新性实验课程体系建设;浙江大学医学院教育改革项目(yxyb20172007)。

摘　　要：凝胶电泳与蛋白质印迹技术是生命医学研究领域的常用技术,但因此项技术步骤较多、操作复杂,学生在实验过程中出现的问题较多,结果不稳定,重复性差。为使学生能熟练且稳定地掌握该技术,对十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)及蛋白质印迹技术进行了长期的实践和探索,通过改进凝胶各成分配比方案,不断完善和优化实验条件,有效增强了凝胶配制的时效性,大大提高了凝胶电泳后续实验的稳定性和成功率。通过优化蛋白印迹过程中抗体的使用浓度,印记杂交结果目标蛋白质条带更加清晰可见,易分析,重复性好。Gel electrophoresis and protein imprinting technology are commonly used in the field of life medicine research, but they have many steps and complicated operations, and students have many problems in the experiment process, with unstable results and poor repeatability.In order to enable students to master this technology skillfully and stably, long-term practice and exploration have been carried out on sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and protein imprinting technology. By improving the formula of gel components,constantly improving and optimizing experimental conditions, the timeliness of gel preparation has been effectively enhanced, and the stability and success rate of subsequent gel electrophoresis experiments have been greatly improved. By optimizing the concentration of antibody used in the process of protein imprinting, the target protein bands in the result of imprinting hybridization are more clearly visible, easy to analyze, and have good repeatability.

关 键 词：凝胶电泳 蛋白质印迹技术 成像系统 实验教学 

分 类 号：R-4[医药卫生] G642.423[文化科学—高等教育学]