不同细胞裂解液提取总蛋白在免疫印迹中的效果分析

目的：寻找一种适合乳腺癌细胞MCF-7蛋白提取的裂解液配方用于免疫印迹分析。方法：配制两种不同的细胞裂解液A与B,分别对培养的密度相当的人乳腺癌细胞MCF-7进行总蛋白的提取,Bradford法测蛋白含量,聚丙烯酰胺凝胶电泳（SDS-PAGE）对蛋白进行分离,免疫印迹法检测β-actin和Twist的表达,比较2种裂解液裂解效果。结果：2种细胞裂解液均可满足免疫印迹分析的要求,B液提取的蛋白含量虽比A液高,但A液用于免疫印迹检测β-actin和Twist的效果更佳。结论：免疫印迹分析结果显示：A细胞裂解液（50 mM pH 7.4 Tris-HCl,150 mM NaCl,0.1%SDS,1%NP-40,1 mM EDTA,临用前加入PMSF使终浓度为1mM）可作为提取乳腺癌细胞MCF-7中蛋白的一种理想配方。

THE EFFECT ANALYSIS OF DIFFERENT CELL LYSATE ON TOTAL PROTEIN EXTRACTION IN WESTERN BLOT

Objective：To search a suitable cell lysate for extraction of proteins from human breast cancer cells MCF-7 for Western blot.Methods：The proteins were extracted from human breast cancer cells by two different cell lysates：A and B.Then total proteins were collected and quantified with the method of bradford.The proteins were separated by SDS-PAGE,and the protein of beta-actin and Twist were assessed by Western blot.Results：The beta-actin and twist of all samples can be observed by Western blot.Though the concentration of proteins extracted by cell lysate A were lower than by cell lysate B,the effect of cell lysate A was better than that of cell lysate B used in Western blot.Conclusion：The cell lysate A is more suitable for extracting proteins from human breast cancer cells MCF-7.