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Two-dimensional gel electrophoresis of proteome in human pancreatic juice
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF PANCREATOLOGY
- Issue:
- 5
- DOI:
- 10.3760/cma.j.issn.1674-1935.2006.05.005
- Key Word:
- 胰液;蛋白质组;双向电泳
Abstract： 目的 建立优化的用于胰液蛋白质组研究的双向电泳方法.方法 经内镜逆行胰胆管造影(ERCP)术收集纯胰液标本5例,分别进行多次双向电泳,比较不同标本保存时间、制备方法、泡胀液组成、固相pH胶条、等电聚焦条件及染色方案等因素对双向电泳图谱质量的影响.结果 胰液标本在-80℃保存4个月做双向电泳对图谱没有明显影响.根据图谱质量比较,三氯乙酸/丙酮沉淀继以含硫脲的泡胀液复溶是优化的标本制备方法;应用窄范围固相pH胶条可以在增加标本上样量的同时改善分辨率;银染和兼容质谱的银染方案所获得的图谱同样具有良好的重复性和分辨率;不同病理状态的胰液标本的双向电泳图谱表现出一定程度的相似性.结论 标本制备是胰液双向电泳的关键,优化的方法可以获得重复性好、分辨率高的双向电泳图谱,为不同病理状态下胰液蛋白质组学差异的研究奠定基础.
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