Establishment and optimization of two-dimensional electrophoresis technique in hydatid fluid proteome of Echinococcus granulosus

The aim of this study was to establish and optimize the two-dimensional electrophoresis (2-de) technology for hydatid fluid proteome of the echinococcus granulosus,and obtain 2-de map of hydatid fluid proteome. total proteins of hydatid fluid were extracted by lyophilization. a series of important factors,such as sample preparation,protein quantities,ph range of immobilized ph gradient (ipg) strip and extraction methods,were optimized to improve the resolution and repeatability. two-dimensional electrophoresis maps were analyzed after staining. the 2-de profiles with high resolution and good repeatability were obtained,when the hydatid fluid dealt with readypreptm 2-d cleanup kit were analyzed with the established 2-de using 400μg of quantitative loading and ipg strips ph7-10. we identified 30 protein spots using pdquest 8.0 2d analysis software. the molecular weight of most of these proteins ranged from 43 to 97kda and the isoelectric points of these proteins ranged from 5 to 9. an optimized 2-de system is set up successfully in this study,electrophoresis pattern of which shows a high resolution and good repeatability,and can be used for the study of e. granulosus proteomics effectively. © 2014 zoological society of pakistan.