人血清中小分子蛋白质分离电泳方法改进

目的建立一种灵敏易行分离分子量〈10000 Da低丰度蛋白质的电泳方法。方法用Hitrap Blue层析柱去除血清中白蛋白,用U9对血清样品进行变性处理,采用改进后的方法（0.1%甲醇,0.5%三氯乙酸,0.1%考马斯亮蓝G250）进行染色。结果经去除血清中的白蛋白、对样品进行变性处理及改进染色方法后,分辨率和灵敏度明显提高,可有效分离10000 Da以下的小分子蛋白质。结论改进后的电泳方法,不仅简便灵敏,而且还有利于蛋白质的洗脱回收。

Separation of micro molecular protein in human serum with improved Tricine - SDS - PAGE system

Objective To separate less abundant proteins less than(10 000) Da with an effective Tricine-SDS-PAGE system.Methods The human serum albumin was removed by Hitrap Blue column,and serum was dealed with U9.Then gel was stained by the improved method(0.1% carbinol,0.5% TCA,0.1%Coomassie briliant blue G250).Results The discovery and detection of less abundant proteins less that(10 000) Da were improved obviously by removal of human serum albumin,serum treated with U9 and improvement of staining.Conclusion The improved Tricine-SDS-PAGE system is not only effective to resolve these less abundant proteins less than(10 000 Da,)but also favourable to recover these proteins from gel.