两种新型聚丙烯酰胺电泳胶电泳特性及保质期研究  

Study on Electrophoresis Characteristics and Shelf Life of Two New Polyacrylamide Electrophoresis Gels

作　　者：李戈辉 吴玮[1,2] 邸铭洋 王长城 杨桂朋 丁海兵[1,2] LI Ge-Hui;WU Wei;DI Ming-Yang;WANG Chang-Cheng;YANG Gui-Peng;DING Hai-Bing(Key Laboratory of Marine Chemistry Theory and Technology,Ministry of Education,Ocean University of China,Qingdao 266100,China;College of Chemistry and Chemical Engineering,Ocean University of China,Qingdao 266100,China;Dycent Biotech(Shanghai)Co.,Ltd,Shanghai 201201,China)

机构地区：[1]中国海洋大学海洋化学理论与工程技术教育部重点实验室,山东青岛266100 [2]中国海洋大学化学化工学院,山东青岛266100 [3]迪申生物技术(上海有限公司),上海201201

出　　处：《中国海洋大学学报（自然科学版）》2021年第1期76-85,共10页Periodical of Ocean University of China

基　　金：山东省重点研究发展计划项目(2018GSF117044);中国海洋大学横向课题项目(20110688)资助。

摘　　要：聚丙烯酰胺凝胶电泳(PAGE)是分离、纯化蛋白质的常用技术。传统的基于Laemmli方法(Laemmli胶)制备的十二烷基磺酸钠聚丙烯酰胺电泳胶(SDS-PAGE)能够基于蛋白质分子量的不同分离蛋白质,蛋白质分离性能良好,但存在电泳时间长、保质期短等缺点,其应用有一定的局限性。本研究用三异丙醇胺、三乙醇胺等成分代替Laemmli胶中的三羟甲基氨基甲烷的盐酸盐(Tris-HCl)作为缓冲物质,获得I型和II型两种新型电泳胶。电泳实验的结果表明这两种新型电泳胶保持了Laemmli电泳胶优良的蛋白质分离性能,并能够耐受更高的电压,具有更快的电泳速度。加速寿命实验的结果表明这两种新型电泳胶具有比Laemmli胶和现在流行的商业预制胶更长的保质期。新型I电泳胶比新型II电泳胶分离性能更好,保质期更长,在37℃可储存15天,相当于在4℃的保质期15个月。Polycarylamide gel electrophoresis(PAGE)is a common technique for protein separation and purification.Sodium dodecyl sulphate-polyacrylamide gelelectrophoresis(SDS-PAGE)based on traditional Laemmli method(Laemmli gel)can separate proteins according to their molecular weight.SDS-PAGE has good efficiency for separating proteins.However,due to the disadvantages of its long electrophoresis time and short shelf life,the application of SDS-PAGE is limited.In this study,triisopropanolamine and triethanolamine were applied to replace Tris(hydroxymethyl)aminomethane(Tris-HCl)in Laemmli gel as buffer reagent,and two new kinds of electrophoresis gels I and II were obtained.The electrophoresis experiments showed that the two new kinds of electrophoresis gel maintained excellent separation performance as Laemmli gel.They can also withstand higher voltage(350 V)with faster electrophoresis speed.Among them,the new I electrophoresis gel has the best performance on standard protein separation at 350 V,and the average fitting degree of the standard curve is not less than 0.9900.The results of accelerated life test indicated that the two new kinds of gel has longer shelf life than Laemmli gel and popular prefabricated commercial gel.The new kind gel I has longer shelf life than the new kind gel II.At 37℃,its shelf life is 15 days,equivalent to 15 months at 4℃.

关 键 词：聚丙烯酰胺凝胶电泳 蛋白质分离 三异丙醇胺 三乙醇胺 电泳胶保质期 

分 类 号：Q-331[生物学—生物化学] Q503