蛋白质组分析中双向聚丙烯酰胺电泳技术的初步建立与优化  

Preliminary establishment and optimization of two dimensional gel electrophoresis(2 DE) for the proteome analysis

作　　者：万晶宏[1,2,3,4] 钱小红[1,2,3,4] 郭尧君 邢桂春[1,2,3,4] 胡志远[1,2,3,4] 邱宗荫 贺福初[1,2,3,4] 

机构地区：[1]军事医学科学院放射医学研究所 [2]军事医学科学院国家生物医学分析中心 [3]中国科学院生物物理研究所 [4]重庆医科大学

出　　处：《军事医学科学院院刊》1998年第4期297-300,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家杰出青年科学基金

摘　　要：目的：初步建立并优化用于蛋白质组分析的双向电泳技术，提高其分辨率及重复性。方法：以ＨｅｐＧ２细胞为例，对以固相ｐＨ梯度为第一向的双向电泳的关键因素与环节，如样品处理、电泳参数和凝胶浓度进行一系列的优化。结果与结论：本研究采用增溶、排异的裂解法处理样品，选择适中的样品量，用预制固相ｐＨ梯度——ＩＰＧ胶条（ｐＨ＝３～１０）第进行第一向等电聚焦，并选用１２％～１４％的梯度胶进行第二向ＳＤＳ分离。Objective: To establish and optimize the two dimensional gel electrophoresis (2 DE) for the proteome analysis, and to improve the resolution and reproducibility. Methods: A series of important factors, such as sample preparation, electrophoresis parameters and gel concentration were optimized with the proteins of HepG2 cell as the example. Results and Conclusion: In present study, adopting the lysis solution which increases sample solubility and eliminates the impurity, choosing appropriate sample amount, using pre cast IPG dry strip and casting 12 14% gradient gel, the 2 DE patterns with good quality have been obtained.

关 键 词：蛋白质组 双向电泳 固相PH梯度 PAGE 

分 类 号：Q51[生物学—生物化学]