高等学校化学学报

高等学校化学学报

• 研究论文 • 上一篇    下一篇

液相等电聚焦结合双向凝胶电泳分离碱性蛋白

钟华, 张扬, 樊惠芝, 杨芃原   

  1. 复旦大学生物医学研究院, 蛋白质组学研究中心, 上海 200433
  • 收稿日期:2006-04-10 修回日期:1900-01-01 出版日期:2006-10-10 发布日期:2006-10-10
  • 通讯作者: 杨芃原

Combination of Liquid Isoelectric Focusing and 2-DE Applied to the Analysis of Alkaline Proteins

ZHONG Hua, ZHANG Yang, FAN Hui-Zhi, YANG Peng-Yuan
   

  1. Proteome Center, Academy of Biomedicine, Fudan University, Shanghai 200433, China
  • Received:2006-04-10 Revised:1900-01-01 Online:2006-10-10 Published:2006-10-10
  • Contact: YANG Peng-Yuan

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被引次数

9

摘要： 在蛋白组学研究中, 经典的双向凝胶电泳法(2-DE)对碱性蛋白及低丰度蛋白的分离存在技术障碍, 但预分离技术的应用可弥补其缺陷. 液相等电聚焦可有效地分离富集复杂蛋白样品. 碱性胶条用于2-DE可极大地提高蛋白上样量和凝胶分辨率. 将上述两种技术相结合用于碱性蛋白质和低丰度蛋白质的分离鉴定, 可使碱端区域双向凝胶图谱质量显著提高, 蛋白点更清晰且点数增多, 质谱鉴定确信度提高, 碱性蛋白和低丰度蛋白质谱鉴定成功率提高, 对于蛋白组学研究具有一定的意义.

关键词: 液相等电聚焦, 双向凝胶电泳, 碱性蛋白

Abstract: Alkaline proteins which represent a large part of proteome, remain a great challenge for conventional 2-DE. Fractionation methods can offset the weakness of 2-DE for the analysis of these proteins. In this work, liquid isoelectric focusing(LIEF) and two-dimensional electrophoresis(2-DE) were conbined for the analysis of alkaline proteins from rat liver. Under this strategy, the resolution of alkaline proteins on 2-D gel and reliability of proteins identification by MS were greatly improved, and more low-abundance and highly alkaline proteins were successfully identified compared to 2-DE without LIEF.

Key words: Liquid isoelectric focusing, Two-dimensional electrophoresis, Alkaline protein

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