双向凝胶电泳分析脾脏蛋白组学的方法研究  

Establishment of two-dimensional polyacrylamide gel electrophoresis techniques for analysis of spleen tissues

作　　者：邱芳华[1] 饶卫农[2] 周强[3] 陈求刚[4] 肖洪广[5] 

机构地区：[1]广州医学院研究生课程进修班,广东广州510182 [2]广州市中医院检验科,广东广州510182 [3]广州医学院附属第二医院检验科,广东广州510260 [4]中山大学附属第一医院检验医学部,广东广州510080 [5]广州医学院附属第一医院检验科,广东广州510080

出　　处：《中国热带医学》2011年第1期10-12,共3页China Tropical Medicine

基　　金：国家自然基金面上项目(30871033)

摘　　要：目的优化双向凝胶电泳的实验步骤,总结一套适用脾脏组织的双向凝胶电泳技术。方法对双向电泳实验中的关键环节:蛋白沉淀方法;胶条PH的选择;聚焦条件等进行研究与优化,考马斯亮蓝染色后进行凝胶图像比较。结果 Cleaning up沉淀蛋白,聚焦电压达到11 000伏小时(110k vhs),可以得到多达1000个蛋白点的2D gel(two-dimensional polyacrylamide gel)。结论建立了脾脏双向电泳的具体方法,得到分辨率高重复性好的2D gel,为脾脏的蛋白质组学研究奠定了基础。Aim To establish a set of two--dimensional polyacrylamide gel electrophoresis （2- DE） techniques for proteomic analysis of spleen tissues. Methods Different methods or condition for key procedures of 2- DE were adjusted and optimized, including sample preparation, choice of IPG PH, condition of iso-eleetronie focusing （IEF）. Comparison of gel eleetrophoresis maps after coomassie staining was carried out. Results High-resolution results with repetitiveness were obtained at condition below：7mmol/L urea,2 mol/L thisurea,4%CHAPS, 100 mmol/L DTT,0.5% （pH 3-10） IPG buffer, 1 mmol/L PMSF as lysis buffer,Cleaning up kit to precipitate protein,IEF to llOk vhs. Conclusion The modified method provided 2-D results with high resolution and repetitiveness, which pave the way for the proteomie analysis of spleen tissues.

关 键 词：脾脏 蛋白质组学 双向电泳 

分 类 号：R521.1[医药卫生—内科学]