Author
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Hurkman Ii, William |
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Tanaka, Charlene |
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Submitted to: Book Chapter
Publication Type: Book / Chapter Publication Acceptance Date: 11/6/2006 Publication Date: 9/9/2007 Citation: Hurkman, W.J., Tanaka, C.K. 2007. High-Resolution Two-Dimensional Gel Electrophoresis: A Cornerstone of Plant Proteomics. In: Samaj, J and Thelen, J (eds.). Plant Proteomics. Berlin, Germany: Springer-Verlag. p. 14-28. Interpretive Summary: The past two decades has seen the introduction of large-scale investigation into the characterization of biological systems. Genomic analysis, in particular assembly of complete genomes and compilation of transcription products, has provided an invaluable foundation for this research. However, the translation of this wealth of genomic data into functional analysis of cellular processes will be in large part dependent upon characterization of the proteome it encodes. Similar to genomic studies, proteomic research has seen the introduction and refinement of a variety of techniques suitable to large-scale analysis. One of the most important of these techniques is high-resolution two-dimensional gel electrophoresis (2-DE) that, in combination with mass spectrometry, provides a central tool for identification of specific gene products and characterization of proteome-wide profiles. This review focuses on protein extraction, solubilization, and detection, key steps to high-resolution 2-DE and maximal proteome coverage and concludes with a brief discussion of comparative proteomics, the quantitative comparison of protein populations for which high-resolution 2-D gels are unrivaled. Technical Abstract: The past two decades has seen the introduction of large-scale investigation into the characterization of biological systems. Genomic analysis, in particular assembly of complete genomes and compilation of transcription products, has provided an invaluable foundation for this research. However, the translation of this wealth of genomic data into functional analysis of cellular processes will be in large part dependent upon characterization of the proteome it encodes. Similar to genomic studies, proteomic research has seen the introduction and refinement of a variety of techniques suitable to large-scale analysis. One of the most important of these techniques is high-resolution two-dimensional gel electrophoresis that, in combination with mass spectrometry, provides a central tool for identification of specific gene products and characterization of proteome-wide profiles. This review focuses on protein extraction, solubilization, and detection, key steps to high-resolution 2-DE and maximal proteome coverage and concludes with a brief discussion of comparative proteomics, the quantitative comparison of protein populations for which high-resolution 2-D gels are unrivaled. |
