摘要
采用固相pH梯度等电聚焦为第一向,SDS PAGE垂直电泳为第二向建立了大肠癌双向电泳分离技术实验条件,对样品的处理、水化、等电聚焦、凝胶平衡等步骤进行了优化,成功地获得了大肠癌分辨率高、重复性好的双向电泳图谱.癌组织样品经3次重复实验共获得蛋白质斑点数1186±46个,蛋白质斑点位置在IEF方向平均偏差为1.67±0.29mm,在SDS PAGE方向为1.41±0.16mm,蛋白质表达量的相对标准差为6.67%±2.25%.经ImageMaster2DElite软件初步分析后发现了一些差异表达的蛋白质.
2DE technique for colorectal carcinoma proteomic research,including the conditions for sample preparation,rehydration,isoelectric focusing,equilibration and other steps are established and improved,a high resolution and reproducible 2DE image is successfully obtained.In three different experiments the total number of protein spots is 1 186±46,the average deviations for protein position in IEF direction is 1.67±0.29 mm and 1.41±0.16 mm in SDSPAGE direction, the relative standard deviations for protein volume is 6.67 %±2.25 %.Some spots show different express after preliminary analysis by ImageMaster 2D Elite software.
出处
《生命科学研究》
CAS
CSCD
2003年第3期214-218,共5页
Life Science Research
关键词
大肠癌
双向电泳
分离条件
固相PH梯度
蛋白质组
immobilized pH gradient
proteomic
two-dimensional gel electrophoresis
colorectal carcinoma
