双向电泳分离大鼠子宫组织提取物的最佳条件摸索

Establishment and Optimization of Two-Dimensional Electrophoresis System for Separating Extract of Rat Uterus

目的:建立并优化大鼠子宫组织提取物的双向电泳分离条件。方法:机械匀浆法制备子宫组织总蛋白提取物,采用不同方法处理样品及不同pH范围的固相梯度胶条进行双向电泳分析。结果:经丙酮沉淀处理的提取物在pH 5～8范围的固相梯度胶条进行双向电泳分离,可获得分辨率清晰和重复性较好的二维图谱。特别是在高分子量蛋白区域得到独立清晰的蛋白质斑点。结论:通过建立并优化双向电泳分离条件,为进一步分析生理与病理状态的子宫蛋白成分提供了技术支持。

Objective： To establish and optimize the two - dimensional electrophoresis for separating extract of rat uterus. Methods： Total proteins of the rat uterus were extracted by mechanical homogenization, treated by different methods and separated by immobilized pH gradient （IPG） strips prior to SDS polyacrylamide gel electrophoresis. Results： The two - dimension- al collection of illustrative plate with better resolution and repetition was achieved when the rat uterus precipitated with TCA - acetone were analyzed with the established two - dimensional electrophoresis using IPG strips pH 5 - 8. Meanwhile, several independent and explicit protein spots were separated in the area of high - molecular - mass proteins. Conclusion： Based on successfully establishing the two - dimensional electrophoresis, it would be helpful to identify the protein constituent of the uterus in physiological and pathological conditions.