聚丙烯酰胺凝胶制板少量分离胶预先快凝防漏方法  

Method of Using Small Amount of Separation Gel to Set Leak Prevention on Polyacrylamide Gel Plate

作　　者：王家俊 朱静怡 朱茵 胡兰婷 宣佳雯 陆思敏 赵起超 谢恬[1,3] 陈功星 WANG Jiajun;ZHU Jingyi;ZHU Yin;HU Lanting;XUAN Jiawen;LU Simin;ZHAO Qichao;XIE Tian;CHEN Gongxing(Division of Health Sciences,Hangzhou Normal University,Hangzhou 311121,China;School of Pharmacy,Hangzhou Normal University,Hangzhou 311121,China;Engineering Laboratory of Development and Application of Traditional Chinese Medicine from Zhejiang Province,Hangzhou 311121,China)

机构地区：[1]杭州师范大学医学部医学院,杭州311121 [2]杭州师范大学医学部药学院,杭州311121 [3]浙产中药材资源开发与应用浙江省工程实验室,杭州311121

出　　处：《实验科学与技术》2022年第6期18-22,共5页Experiment Science and Technology

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);浙江省中医药科技计划项目(2021ZB185)。

摘　　要：利用聚丙烯酰胺凝胶制板新旧玻璃板和密封垫,预先快凝少量分离胶堵漏方法(DIY)各30次制胶效果比较,并用DIY制胶进行了Jurkat和A549细胞株蛋白样本电泳、转膜染色和Actin抗体反应实验,发现新玻璃板和密封垫制胶优于部分和全旧的器具(P <0.05),而旧玻璃板和密封垫的DIY制胶与新器具没有区别(P>0.05),且在后期的蛋白质样本分析中发现凝胶及其转膜蛋白染色正常,两种细胞的Actin蛋白表达灰度值显示没有统计学差异(P>0.05)。因此DIY方法延长了玻璃板和密封垫的有效使用期,节约了经费,成为一种简便实用的聚丙烯酰胺凝胶板制备方法。We studied the polyacrylamide gel-casting new and old glass plates and gasket in the preparation of small amount of separation gel to seal on polyacrylamide gel plate(Do it yourself,DIY) and compared the effects thirty times each of them.After that the DIY gel was used to electrophorese the protein samples of Jurkat and A549 cell lines,then strain the transformed PVDF with Coomassie blue dye and detect Actin antibody reaction with Western blot.The results showed that the success rate of gel-casting new plate and gasket was the best to whole or a part of the old those with statistically significant difference(P 0.05).As same time the DIY gel was used in the subsequent experiments.It was normal that the protein samples in electrophoresis gel and PVDF Coomassie blue staining.The integrated intensities of Actin expression of two cell line were the same(P >0.05).So by the DIY method could prolong the service life of gelcasting plate and gasket with funds retrenched.It had become a simple and practical method to prepare polyacrylamide gel plate.

关 键 词：聚丙烯酰胺 凝胶 制板 防漏 电泳 

分 类 号：R446.1[医药卫生—诊断学]