大鼠皮质双向电泳样品制备方法的优化  

Optimizing Preparation of Rat Cortex Two-dimensional Electrophoresis Sample

作　　者：张永杰[1] 徐雅飞[1] 王群[1] 王建枝[1] 

机构地区：[1]华中科技大学同济医学院基础医学院病理生理学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2006年第1期121-123,共3页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30328007;30170221;30100057)

摘　　要：目的优化大鼠皮质双向电泳样品制备方法,初步建立双向电泳条件,提高电泳分辨率和重复性。方法分别应用三氯乙酸/丙酮沉淀法、超速离心法及改良超速离心法制备电泳样品,并比较各处理方法对电泳的影响。改良超速离心法即将混合了匀浆液Ⅱ的匀浆物4℃放置过夜后再进行后续处理。结果改良超速离心法所制备的电泳样品,不仅可以减少蛋白的降解,还可以增加蛋白的溶解性,从而获得分辨率较高、重复性较好的双向电泳图谱。结论改良超速离心法更适于制备来源于中枢神经系统双向电泳样品。Objective To optimize rat cortex sample preparation, primarily establishing parameters of two-dimensional eleetrophoresis, and improving its resolution and reprodueibility. Methods By using TCA/Acetone-precipitation. uhracentrifugallon and modified uhracentrifugation, rat cortex samples were prepared and the effects of different protein extraction methods on the electrophoresis resuhs were compared. Results Modified uhracentrifugation not only decreased protcin decomposition. but also increased protein solubility, therefore obtaining the two dimensional electrophoresis speetrum with higher resolution and belier reproducibility. Conclusion Modified uhraeentrifugation is more suitable for preparing the sample of eentral nervous system.

关 键 词：皮质 双向电泳 三氯乙酸/丙酮沉淀法 超速离心法 

分 类 号：R745.7[医药卫生—神经病学与精神病学] Q513[医药卫生—临床医学]