Protein isolation and second-dimension electrophoretic separation.

Bernhard Schlesier; Hans-Peter Mock

doi:10.1385/1-59745-003-0:381

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Protein isolation and second-dimension electrophoretic separation.

Bernhard Schlesier·Hans-Peter Mock

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Abstract

Protocols for the proteome analysis of Arabidopsis tissues based on 2 D gel electrophoresis are given for leaves and roots as well as for seeds. The protocols contain a detailed description of the sample preparation step and of the solubilization of proteins. We then describe our protocols for the isoelectric focusing, the transfer of the strips from the first to the second dimension, SDS-PAGE as the second dimension of electrophoretic separation, staining of the gels using colloidal Coomassie blue, and finally drying of the gels between cellophane sheets. Notes contain additional background information on critical steps. Our protocols allow the analysis of leaves, roots, or seeds from different Arabidopsis ecotypes, from mutants or from plants subjected to various environmental conditions.

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Arabidopsis Electrophoresis, Gel, Two-Dimensional Electrophoresis, Polyacrylamide Gel Isoelectric Focusing Plant Leaves Plant Proteins Plant Roots Proteomics Rosaniline Dyes Journal Article Research Support, Non-U.S. Gov't

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Bernhard Schlesier, Hans-Peter Mock. 2006. Protein isolation and second-dimension electrophoretic separation.. https://doi.org/10.1385/1-59745-003-0%3A381

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