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目的:建立结肠癌13cm和24cm非线性分离系统的2-D图谱,分析比较两者的分辨率。方法:提取结肠癌总蛋白,用pH3-10非线性干胶条对样品进行等电聚焦分离,并分别使用13cm和24cm电泳系统进行双向电泳,考马斯亮蓝G250染色,图像分析,比较对比两组2-D图谱,量化分析两种系统的分辨率差异。结果:在等点电3-10,分子量20-170kD范围内分别分离得到蛋白质斑点873个(13cm电泳系统)和1349个(24cm电泳系统)。对于24cm电泳系统,1mg蛋白质上样量的电泳图谱清晰,分辨率较好。结论:成功建立了高分辨率、简便易控的结肠癌蛋白质组双向电泳技术平台。
Objective: To establish a 2-D map of 13cm and 24cm non-linear separation system of colon cancer, analyze and compare the resolution of the two. Methods: Total colon cancer protein was extracted. The samples were separated by isoelectric focusing using pH3-10 non-linear dry strip. Two-dimensional gel electrophoresis was performed on 13cm and 24cm electrophoresis systems respectively. Coomassie brilliant blue G250 staining and image analysis were performed. 2-D spectra, quantitative analysis of the resolution difference between the two systems. Results: 873 protein spots (13cm electrophoresis system) and 1349 (24cm electrophoresis system) were isolated from the isoelectric point of 3-10 and the molecular weight of 20-170kD respectively. For 24cm electrophoresis system, 1mg protein sample electrophoresis clear, better resolution. Conclusion: A high-resolution, simple and easy-to-use protocol for two-dimensional electrophoresis of proteome of colon cancer has been successfully established.