牛乳碱性蛋白二维凝胶电泳条件的优化

Optimization of Two-dimensional Gel Electrophoresis for Separating Basic Protein in Bovine Milk

为了在二维凝胶电泳(2-DE)中有效分离牛乳中的碱性蛋白,本试验在等电聚焦上样缓冲液中加入异丙醇和甘油,采用杯上样方式,比较了三丁基磷(TBP)和二硫苏糖醇(DTT)两种还原剂对碱性区域蛋白的分离效果。结果发现,等电聚焦上样缓冲液以TBP为还原剂的碱性蛋白分离图谱水平条纹少,优于以DTT为还原剂图谱。表明等电聚焦上样缓冲液以TBP为还原剂,采用杯上样的方法可以改善牛乳中碱性蛋白的分离效果。

Two-dimensional gel electrophoresis （2-DE）was used to separate the high resolution of basic protein from bovine milk. In this study, dithiothreitol（DTT） was replaced by tributyl phosphine （TBP）as the reducing agent in the sample solution for the first-dimensional isoelectric focusing. The sample was cup-loaded at the anode. The results showed that, the combined use of TBP and anodic cup-loading improves protein solubility during isoelectric focusing, which results in higher resolution. The findings may provide valuable information for the separation of basic protein.