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Sequence information and preparation of resolved by; two‐dimensional gel electrophoresis: no longer just spots on gels
Author(s) -
Guy Charles L.
Publication year - 1989
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1989.tb05488.x
Subject(s) - isoelectric point , isoelectric focusing , gel electrophoresis , two dimensional gel electrophoresis , polyacrylamide gel electrophoresis , electrophoresis , molecular mass , chemistry , chromatography , biochemistry , proteomics , enzyme , gene
In 1975 O'Farrell described, in detail, a procedure to separate proteins by polyacrylamide gel electrophoresis in two dimensions. This powerful new technique relied on two characteristics of proteins: charge and molecular mass. In the first dimension, proteins were separated on the basis of net charge in a pH gradient by isoelectric focusing, and in the second dimension the proteins were further fractionated exclusively on the basis of their molecular mass by SDS gel electrophoresis. Since two‐dimensional gel electrophoresis (2DGE) has a resolving power of at least 20 fold greater than one‐dimensional electrophoresis, it has found wide spread application in modern biological research. However, beyond the detection of a given protein, 2DGE provides little additional information about a specific protein other than molecular mass, isoelectric point, and approximate relative abundance. In recent years, the development of new technologies have made it possible to directly obtain sequence information, and produce specific antisera for proteins resolved by 2DGE. These new technological developments serve to further increase the power and utility of 2DGE in the analysis of proteins of importance to plant physiology.