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.
出处
《军事医学科学院院刊》
CSCD
北大核心
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
关键词
蛋白质组
双向电泳
固相PH梯度
PAGE
proteome
two dimensional gel electrophoresis
immobilized pH gradients
HepG 2 cell
