中国组织工程研究与临床康复2009，Vol.13Issue(28)：5440-5443,4.DOI:10.3969/j.issn.1673-8225.2009.28.008
家兔血清蛋白质组双向凝胶电泳技术的建立
Establishment of two-dimensional gelelectrophoresis technique for rabbit serum proteomics
摘要
Abstract
BACKGROUND: Rabbit serum samples are widely used in basic researches, and two-dimensional gelelectrophoresis (2-DE) is the most classic technique for protein separation. Therefore, it is of great significance to establish a stable technique system of 2-DE for rabbit serum.OBJECTIVE: To establish a 2-DE technique system for rabbit serum protein separation. DESIGN, TIME AND SETTING: A single sample observational experiment was performed at Tianjin Key Laboratory of Biomarkers for Occupational and Environmental Hazard of Chinese People's Armed Police Forces Medical College from June to July in 2008. MATERIALS: Six healthy rabbits were provided by the Animal Experimental Center of Tangshan Vocational Technical College METHODS: Health rabbit serum was dissolved in rehydration sample loading buffer before and after eliminating high abundance proteins to make proteins in it schizolysis adequately. After reductive alkylation, the samples were loaded into the rehydration tray to undergo passive rehydration for 14 hours. Isoelectric focusing (IEF) electrophoresis was followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After sliver staining, the gel was analyzed by PDQuest7.4. MAIN OUTCOME MEASURES: ①Efficiency of eliminating high abundance proteins. ②Two-dimensional eleotrophoregrams (2-D electrophoregrams)RESULTS: Distinct 2-D eleotrophoregrems were obtained with high resolution and good reproducibility. The removal of high abundance proteins in serum failed to result in better 2-D electrophoregrams.CONCLUSION: We have successfully established a 2-DE technique for rabbit serum proteome, which can lay the foundation for the further study of serum proteomics of diseases.关键词
家兔/血清蛋白质组/双向凝胶电泳分类
医药卫生引用本文复制引用
要瑞莉,张明顺,李宏杰,董淑云,王世鑫,冯桂玲..家兔血清蛋白质组双向凝胶电泳技术的建立[J].中国组织工程研究与临床康复,2009,13(28):5440-5443,4.基金项目
河北省科学技术研究与发展指导计划项目(072761489) （072761489）
a grant from the Science and Technology Research and Development Program of Hebei Province, No. 072761489 （）
