同位素标记结合免疫印迹-化学发光法鉴定小鼠神经元磷酸化蛋白质组  

Identification of Phosphoproteome in Mice Neurons by Isotope-Labeling Technique Combining with Chemiluminescence Western Blotting Method

作　　者：姚富丽[1] 李洪[1] 尹康[2] 何涛[3] 吴民泸[1] 肖斌[1] 

机构地区：[1]泸州医学院基础医学院生物化学教研室,泸州646000 [2]泸州医学院附属医院外总教研室,泸州646000 [3]泸州医学院科研处,泸州646000

出　　处：《生物医学工程学杂志》2007年第4期898-901,共4页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(70071040);泸州医学院科研基金资助项目

摘　　要：创建一套简单、实用、灵敏、高效的分析鉴定磷酸化蛋白质组的优化方案。采用同位素标记、双向电泳、放射自显影等技术建立小鼠神经元磷酸化蛋白质组图谱,再用免疫印迹-持久性化学发光技术选择性地鉴定三种小鼠神经元磷酸化蛋白质——PI3Kr3、MEK1、PKCα。结果显示三种磷酸化蛋白质在双向电泳放射自显影图谱和双向电泳免疫印迹-化学发光图谱上的斑点基本匹配,提示本研究建立的以同位素标记和免疫印迹-化学发光法为核心的分析鉴定磷酸化蛋白质组的一整套优化方案灵敏度高、特异性强。This study was aimed to work out a simple, applicable, sensitive and specific protocol for the identification of phosphoproteome. Isotope-labeling, two-dimensional electrophoresis, autoradiography and so on were used to establish a phosphoproteome map of mice neurons,and then chemiluminescence Western blotting was utilized to detect three phosphoproteins PI3Kr3, MEK1 and PKCa selectively. The results of comparison showed that the blots of PI3Kr3, MEK1 and PKCα on autoradiography map were almost identical with the blots of PI3Kr3, MEK1 and PKCα on chemiluminescence Western blotting maps. So this protocol based on isotope labeling and chemiluminescence Western blotting methods has proven to be sensitive and specific in the identification of phosphoproteome.

关 键 词：同位素标记 双向电泳 免疫印迹 化学发光 磷酸化蛋白质组 

分 类 号：R817[医药卫生—影像医学与核医学]