摘要
目的:建立并优化乙型肝炎肝纤维化组织蛋白质组研究所需的双向凝胶电泳技术,提高其分辨率和重复性.方法:外科手术获取乙型肝炎肝纤维化组织,提取肝组织总蛋白.利用固相pH梯度双向凝胶电泳技术分离总蛋白,考马斯亮蓝染色后分析凝胶图像.对2-DE中的关键因素与环节,如样本处理、上样量、电泳参数等各步条件进行了一系列优化.结果:优化条件后的乙型肝炎肝纤维化组织双向凝胶电泳图谱水平拖尾减少、斑点分辨率改善、斑点检出数量增加(246±33→729±83).结论:该双向凝胶电泳技术可应用于乙型肝炎肝纤维化组织的蛋白质组学研究.
AIM： To establish and optimize the technique of two-dimensional gel electrophoresis （2-DE） for studying the proteomics of liver fibrosis tissue from patients with hepatitis B.
METHODS： Liver fibrosis tissues were obtained by surgical method, and the total proteins were extracted. Immobilized pH gradient isoelectric two-dimensional gel electrophoresis was used to separate the total protein. After coomassie brilliant blue staining, 2-DE maps were analyzed with ImageMaster 2D Platinum Software. A series of conditions such as the sample size, electrophoresis parameters and the way to treat the sample were improved.
RESULTS： After optimization, the horizontal trail was decreased; the resolution of the spot was improved; the number of protein spot was increased from 246 ± 33 to 729 ± 83 in the 2-DE maps.
CONCLUSION： The optimized technique of 2-DE can be introduced to study the proteomics of liver fibrosis tissue from patients with hepatitis B.
出处
《世界华人消化杂志》
CAS
北大核心
2007年第13期1554-1557,共4页
World Chinese Journal of Digestology
基金
贵州省优秀青年科技人才基金资助
No.(2005)0518
贵州省优秀科技教育人才省长专项
No.(2005)223
贵州省卫生厅科技
No(2006)046~~
关键词
乙型肝炎
肝纤维化组织
双向凝胶电泳
Hepatitis B, Liver fibrosis tissue
Two-dimensional gel electrophoresis
