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使用固定化pH梯度管状凝胶的二维凝胶电泳。

Two-dimensional gel electrophoresis using immobilized pH gradient tube gels.

作者信息

Hirano H, Kawasaki H, Sassa H

机构信息

Yokohama City University, Kihara Institute for Biological Research/Graduate School of Integrated Science, Totsuka, Japan.

出版信息

Electrophoresis. 2000 Jan;21(2):440-5. doi: 10.1002/(SICI)1522-2683(20000101)21:2<440::AID-ELPS440>3.0.CO;2-X.

Abstract

An apparatus for the preparation of gels for immobilized pH gradient isoelectric focusing (IPG) in glass tubes was developed. Using this apparatus, the highly reproducible immobilized pH gradient can be formed with Immobilines in polyacrylamide gels, and IPG gels at all possible pH ranges can be easily prepared at low cost. The IPG tube gels in the first dimension in two-dimensional gel electrophoresis was used to separate and identify a number of rice embryo proteins in the proteome analysis. There was no difference in resolution of proteins between the tube gels and the commercially available slab gels; after electrophoresis, however, we could efficiently obtain a larger amount of the purified proteins from the tube gels than from the slab gels.

摘要

开发了一种用于在玻璃管中制备用于固定化pH梯度等电聚焦(IPG)凝胶的装置。使用该装置,可以在聚丙烯酰胺凝胶中用固定化电解质形成高度可重复的固定化pH梯度,并且可以以低成本轻松制备所有可能pH范围内的IPG凝胶。二维凝胶电泳第一维中的IPG管凝胶用于蛋白质组分析中分离和鉴定多种水稻胚蛋白质。管凝胶和市售平板凝胶之间蛋白质的分辨率没有差异;然而,电泳后,我们从管凝胶中比从平板凝胶中能更有效地获得大量纯化蛋白质。

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