| 一种新的双向电泳蛋白样品制备方法 |
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| 引用本文： | 李小兰,谢大兴,肖徽,吴剑宏,姚静,冯永东,陶德定,刘双又,胡俊波,龚建平.一种新的双向电泳蛋白样品制备方法[J].华中科技大学学报(医学版),2006,35(5):662-664. |
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| 作者姓名： | 李小兰 谢大兴 肖徽 吴剑宏 姚静 冯永东 陶德定 刘双又 胡俊波 龚建平 |
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| 作者单位： | 华中科技大学同济医学院附属同济医院肿瘤研究所分子医学中心,武汉,430030 |
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| 基金项目： | 国家重点基础研究发展计划(973计划);国家自然科学基金 |
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| 摘 要： | 目的探讨双向凝胶电泳（2-DE）样品裂解液对蛋白质分离和双向凝胶电泳结果的影响。方法以MOLT-4细胞为例，采用3种溶解性能不同的裂解液提取细胞中的蛋白质组，并分别进行双向聚丙烯酰胺凝胶电泳。结果通过对2-DPAGE图谱的比较分析，发现采用8mol／L Urea，2mol／L Thiourea，4％CHAPS，1％NP-40.4％Triton X-100，65mmol／LDTT，0．5mmol／LPMSF，0．5％pharmalyte3-10，能获得质量较高的2-DE图谱。结论采用高浓度脲、三去污裂解液有利于获得优质的蛋白质样品，从而提高蛋白质提取率和双向电泳分辨率等。
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| 关 键 词： | 蛋白质组 双向电泳 固相pH梯度 MOLT-4细胞 |
| 修稿时间： | 2005年10月28 |
A New Preparation Method for the Protein Sample in Two-dimensional Electrophoresis |
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| Li Xiaolan,Xie Daxing,Xiao Hui et alMolecular Medicine Center,Cancer Research Institute,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan.A New Preparation Method for the Protein Sample in Two-dimensional Electrophoresis[J].Journal of Huazhong University of Science and Technology(Health Sciences),2006,35(5):662-664. |
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| Authors: | Li Xiaolan Xie Daxing Xiao Hui Molecular Medicine Center Cancer Research Institute Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan |
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| Affiliation: | Li Xiaolan,Xie Daxing,Xiao Hui et alMolecular Medicine Center,Cancer Research Institute,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030 |
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| Abstract: | Objective To investigate the effects of the sample lysis solution on protein separation and results about two-dimensional electrophoresis in two-dimensional gel electrophoresis.Methods Three different resolvable solutions were adopted to take out the proteins of MOLT-4 cells as the example,and then the proteins were separated by two dimensional polyacrylamide gel electrophoresis.Results By using 8 mol/L Urea,2 mol/L Thiourea,4 % CHAPS,1 % NP-40, 4 % TritonX-100,65 mmol/L DTT,0.5 mmol/L PMSF,0.5 % pharmalyte 3-10 much better quality map of 2-DE could be obtained.Conclusion Using lysis solution containing high concentration urea and three different detergents can obtain more quality samples,then increase the extraction rate of proteins and resolution about two-dimensional electrophoresis. |
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| Keywords: | proteome two-dimensional gel electrophoresis immobilized pH gradients MOLT-4 cell |
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