【摘要】 目的建立并优化人胰液蛋白质组的双向凝胶电泳分离方法。方法通过比较几种常用的体液处理方法,确定了一种适于人胰液的蛋白质样品处理方法。经过一维等电聚焦和二维SDS-PAGE电泳后,有效分离了人胰液蛋白质组。结果通过优化胰液蛋白质提取、上样量和固相IPG胶条分离范围的选择等步骤,建立了人胰液双向凝胶电泳的方法,并获得了分辨率较高、重复性较好的双向凝胶电泳图谱。结论丙酮沉淀与双向凝胶电泳技术的结合是研究人胰液蛋白质组的一种有效的方法。胰液双向凝胶电泳方法的优化可为今后开展疾病相关的胰液蛋白质组学研究提供技术基础。更多还原

【Abstract】 Objective To establish and optimize the two-dimensional gel electrophoresis separation of proteome in human pancreatic juice.Methods By comparing several commonly used protein extraction methods,a method for the separation of proteome in human pancreatic juice was determined.The proteome in human pancreatic juice was effectively concentrated and separated by isoelectric focusing on first dimension and SDS-PAGE on second dimension.Results By optimizing the extraction of proteins in human pancreatic juice,loading amount and separation scope of IPG strip,the proteins were successfully extracted from human pancreatic juice and were separated by two-dimensional gel electrophoresis with higher resolution and better reproducibility.Conclusion Acetone-precipitation combined with two-dimensional gel electrophoresis is a suitable approach to study the proteome in human pancreatic juice.The optimization of the separation of human pancreatic juice by two-dimensional gel electrophoresis lays a technological foundation for the further investigation of disease-related human pancreatic juice proteomics.更多还原