摘要
目的:建立基于二维液相色谱法的三维分离小胶质细胞蛋白质组的方法。方法:将小胶质细胞裂解后制备全细胞蛋白,在传统的一维HPCF、二维HPRP分离后,增加了三维SDSPAGE电泳,对小胶质细胞实现了三维分离。结果:经过优化后的三维分离,小胶质细胞全蛋白按照pH值由高到低的顺序分为34个组分,其中pH8.5～4.0的部分包含16个组分,每一个组分又包含若干种蛋白质。结论:同传统的二维液相色谱分离方法相比,改进后的三维分离方法对小胶质细胞全蛋白具有更彻底、更好的分离效果,为后续的质谱鉴定提供了分离度更好的样品。
Objective:To develop a three dimensional separation method for separating microglial proteome by three dimensional fraction based on 2D liquid phase chromatographic fractionation(PF-2D). Methods: Microglial lysate was used to prepare whole cell proteome, which was then subjected to three dimensional separation: the first dimensional separation (HPCF), the second dimensional separatim (HPRP) and the third SDS-PAGE electrophoresis.Results: After three dimensional separation,microglial proteome was graded into 34 groups in the descending order of pH decreased. There were 16 groups between pH 8.5 and 4.0,and every group included some proteins.Conclusion: Compared with the traditional 2D liquid phase chromatographic fraction, the modified method is more effective for separating microglial proteome,providing purer protein for mass spectrometric identification.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2005年第6期687-689,共3页
Academic Journal of Second Military Medical University
基金
国家高新技术发展规划("863"计划)课题(2002AA2Z3325).
