Electroblotting of Proteins from 2-D Polyacrylamide Gels
By: Michael J. Dunn2
| Abstract |
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The ability of two-dimensional electrophoresis (2-DE) to separate complex mixtures of proteins, such as represented by cells,
tissues, and even whole organisms, has been recognized for more than 20 yr (1). Using “standard”-format (around 20Ã20 cm) 2-D gels, the method is capable of routinely separating 2000 proteins from whole-cell
and tissue extracts. The resolution capacity can be extended significantly (up to 5000–10,000 proteins) using large-format
(40Ã30 cm) 2-D gels (2). Over the last 20 yr, 2-DE has been used primarily as an analytical tool for the characterization of proteins by their charge
(pI), size (M
r), and relative abundance. Specialized computer software has been developed for the qualitative and quantitative analysis
of 2-DE protein patterns (3), and these systems have been used to construct several comprehensive databases of protein expression in a variety of cell
types and tissues (for examples, see
ref.
4).
Affiliation(s): (2) Department of Cardiothoracic Surgery, National Heart and Lung Institute, Heart Science Centre, Harefield Hospital, Harefield, Middlesex, UK
Book Title: 2-D Proteome Analysis Protocols
Series: Methods in Molecular Biology | Volume: 112 | Pub. Date: Sep-24-1998 | Page Range: 313-318 | DOI: 10.1385/1-59259-584-7:313
Subject: Biochemistry
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