结肠癌蛋白质双向电泳技术的建立

Establishment of a two-dimensional electrophoresis technology on colon carcinoma

目的:建立结肠癌双向电泳技术。方法:利用不同体积的裂斛液提取结肠癌中的蛋白质,并通过不同的蛋白质上样量(1mg,2mg,3mg),不同样品制备方式,进行双向电泳,考马斯亮蓝染色,图谱分析。结果:在等电点3～10,分子量6.5～200ku范围内分离得到蛋白质斑点为,1mg蛋白质上样量为658个蛋白质斑点,2mg为820个,3mg为845个斑点。2mg蛋白质上样量的双向电泳图谱更清晰,分离更好。制备方式2可以满足双相电泳的需要。结论:成功建立了结肠癌蛋白质的双向电泳技术。

Objective：To establish a two-dimensional electrophoresis technology tissue protein of colon carcinoma. Methods： Lysis buffer of different volume was taken to extract tissue proteins of colon carcinoma,and different protein quantities （1mg,2mg,3mg） different preparation ways were taken to establish a two-dimensional electrophoresis. Coomassie brilliant blue was applied to stain protein,and patterns were analyzed. Results：With a molecular mass between 6.5-200ku and isoelectric points （pI） from 3-10,1mg proteins obtained 569 protein spots,2mg 786,and 3mg 835. Pattern of 2mg protein and the sample two were the best. Conclusion：A two-dimensional electrophoresis technology on tissue protein of colon carcinoma has been established successfully.