摘要
目的:优化双向电泳的样品处理方法,建立HepG2细胞蛋白质双向电泳图谱。方法:用含10%胎牛血清的RPM11640培养液培养HepG2细胞,胰酶消化后收集细胞,加入细胞裂解液使细胞充分裂解,冰浴超声离心后取上清液进行双向凝胶电泳,电泳图谱经考马斯亮蓝染色及图像扫描与分析。结果:通过对HepG2细胞蛋白提取液进行双向电泳,在13 cm pH 3~10(L)IPG胶条上可分离到(297±23)个重复性及分辨率均较好的蛋白质点,蛋白斑点的匹配率为83%。结论:HepG2细胞蛋白质组双向电泳条件的优化及图谱的建立,为进一步研究肝癌蛋白质组学奠定了基础。
Objective:To optimize the treatment methods of Two-dimensional Electrophoresis sample and establish the 2-DE profiles of proteome of HepG2 cells.Methods: HepG2 cells were cultured in RPMI 1640 culture medium containing 10% fetal bovine serum,then harvested after digestion by trypsin,and lysed in lysis buffer.After centrifugation,supernatant was separated by two-dimensional gel electrophoresis,stained by Coomassie Brilliant Blue,and then followed by image scanning and analysis.Results:(297±23)protein spots were detected in the 2-DE profiles on 13 cm IPG strip(pH 3~10 L) of HepG2 cells proteome separated by 2-D PAGE.A 83% match rate of protein spots between gels was obtained.Conclusion:The establishment and optimization of two-dimensional gel electrophoresis technique for proteomics of HepG2 cells laid foundation for further research on hepatocellular carcinoma proteomics.
出处
《内蒙古医学杂志》
2011年第5期527-529,F0004,共4页
Inner Mongolia Medical Journal
关键词
肝癌
蛋白质组学
双向电泳
Hepatocellular carcinoma
Proteomics
Two-dimensional gel