胆管癌胆汁蛋白质样品制备和双向电泳条件的优化  

Optimization of Sample Preparation and Two-dimensional Gel Electrophoresis for Cholangiocarcinoma Bile Protein

作　　者：吕济相[1] 王济明[1] 罗诗樵[1] 黄国飞[1] 

机构地区：[1]重庆医科大学附属第一医院肝胆外科,重庆400016

出　　处：《生物技术通报》2009年第8期125-127,133,共4页Biotechnology Bulletin

基　　金：重庆市卫生局2007年度医学科研项目计划(07-01-003)

摘　　要：采用合适的裂解液和沉淀方法,提取胆管癌胆汁中的蛋白质,获得分辨率高、重复性好的蛋白质双向电泳图谱。通过不同裂解液和蛋白质沉淀方法提取胆汁蛋白的效果比较,设计了不同的样品制备方法,并且对双向电泳(2-DE)的条件进行优化。结果显示,试验确定了适合胆管癌胆汁的裂解液配方(LSⅣ),丙酮沉淀的蛋白分布完整,沉淀效率相对较高。高伏时、长时间的等电聚焦可以获得高分辨率、重复性好的蛋白质双向电泳图谱。因此,本方法可以应用到胆管癌胆汁蛋白的提取,也可以对其他体液蛋白质样品的制备和双向电泳提供借鉴。It was o obtain the high resolution and better reproducibility two-dimensional gel electrophoresis（2-DE）of protein with extracted cholangiocarcinoma bile protein by suitable lysate and sediment methods. The effect of bile total protein extracted by protein lysate and sediment methods was compared to design different method to prepare the protein sample and to optimize 2-DE. Results showed that the suitable lysate prescription IV （ LS IV） for cholangiocarcinoma bile were set according to the complete protein distribu- tion of acetone sediment and the higher sediment rate. High-voltage and long-running time in IEF could give rise to high-resolution and preferable repro-ducibility 2-DE image. Thus, it proved that this method could be applied to extract the protein in cholangiocarcinoma bile and prepare the other body fluid protein sample and 2-DE studies.

关 键 词：胆管癌 样品制备 双向电泳 

分 类 号：R735.8[医药卫生—肿瘤]