文献类型： 外文期刊

第一作者： Fang Qinmei

作者： Fang Qinmei;Sun Wei;Jiao Liyan;Qian Xiaohong;Jiang Ying;Xia Chun;He Fuchu

作者机构：

关键词： hepatoma carcinoma cell;cell collection;protein extraction;two-dimensional differential in-gel electrophoresis;system evaluation

期刊名称：CHINESE JOURNAL OF ANALYTICAL CHEMISTRY （ 影响因子：1.134； 五年影响因子：0.909 ）

ISSN： 0253-3820

年卷期： 2006 年 34 卷 11 期

页码：

收录情况： SCI

摘要： The research on comparison of the difference of hepatoma carcinoma cell and hepatocytes differential proteomics two-dimensional differential in-gel electrophoresis (2D-DIGE) technique needs to have a labeling system evaluation on its protein sample for success of DIGE experiment. Celluar proteomics research should collect most proteins in cells, so mechanical erasion, trypsinization and PBSE digestion were tested to collect HepG2 cell, the result shows that the PBSE digestion can keep the integrated cell morphous mostly and get the topmost protein yield. Then sodium dodecylsulfonate polyacrylaminde gel chromatography ( SDSPAGE) and two dimensional electoporesis (2DE) of the protein sample labeled with 3 kinds of fluorescents run. The scanning image of SDS-PAGE and 2DE gel of HepG2 cell protein were analyszed by ImageQuant and DeCyder 7. 0 in differential in-gel analysis ( DIA) and biological variation analysis ( BVA) models, repectively. The Image quant result displays that the sample can mutually compatible with the dye and the protein quant have a better linear relationship with the fluorescence intensity. The DIA result shows that the quantitation of protein labeled with Cy3 and Cy5 is mainly equal except those low molecular weight extremely acidic and basic protein tending to migration on 2DE gel, which may result in the error of protein quantitation analysis, but the inversely labeling strategy can correct the quantitation error. On the whole, system evaluation of DIGE technique shows that test labelinp and protein quantitation are successful, which can provide advantageous references for carcinoma cell DIGE experiment, and hints that the protein can be analyzed by 2D-DIGE. The system evaluation method firstly verify the reliabity of DIGE system and give an error range of quantitation,which offers a reference to DIGE platform.

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