The aim of Plant Proteomics: Methods and Protocols is to present up-- date methods and protocols used by recognized scientists in the world of plant proteomics. If this world was a very small one twenty-five years ago when the first papers were published, it has since experienced exponential growth, and in most countries around the world there are laboratories working on plant proteomics. Two-dimensional gel electrophoresis is still the basic method used, but it has been improved greatly with IPG in the first dimension (Chapter 13) and with new detection methods with fluorochromes (Chapters 14 and 15). Signi- cant progress has been achieved in protein extraction, which is particularly difficult with plant tissues containing phenols, proteases, and other secondary metabolites that interfere with proteins. Standard procedures have been op- mized (Chapters 1 and 2) for peculiar tissues (Chapters 3, 4, and 5) and cellular compartments (Chapters 6 to 10). These methods rely on improvements made in the solubilization of proteins from membranes (Chapters 11 and 12). Mass spectrometry was a revolution that permitted the high throughput identifi- tion of proteins separated by 2D gels (Chapters 19 and 20) but also from blue native 1D gels (Chapters 27 and 28) despite the fact that Edman sequencing can still be useful (Chapter 18). Associated with other techniques such as 2DLC or LC of intact proteins, mass spectrometry also permits the identification of polypeptides from complexes (Chapters 21 and 22).
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The aim of Plant Proteomics: Methods and Protocols is to present up-- date methods and protocols used by recognized scientists in the world of plant proteomics. If this world was a very small one twenty-five years ago when the first papers were published, it has since experienced exponential growth, and in most countries around the world there are laboratories working on plant proteomics. Two-dimensional gel electrophoresis is still the basic method used, but it has been improved greatly with IPG in the first dimension (Chapter 13) and with new detection methods with fluorochromes (Chapters 14 and 15). Signi- cant progress has been achieved in protein extraction, which is particularly difficult with plant tissues containing phenols, proteases, and other secondary metabolites that interfere with proteins. Standard procedures have been op- mized (Chapters 1 and 2) for peculiar tissues (Chapters 3, 4, and 5) and cellular compartments (Chapters 6 to 10). These methods rely on improvements made in the solubilization of proteins from membranes (Chapters 11 and 12). Mass spectrometry was a revolution that permitted the high throughput identifi- tion of proteins separated by 2D gels (Chapters 19 and 20) but also from blue native 1D gels (Chapters 27 and 28) despite the fact that Edman sequencing can still be useful (Chapter 18). Associated with other techniques such as 2DLC or LC of intact proteins, mass spectrometry also permits the identification of polypeptides from complexes (Chapters 21 and 22).
Plant Proteomics: Methods and Protocols provides up-to-date methods and protocols for plant proteomics, each written by expert researchers from leading laboratories. Experimental methodologies are explained in clear and complete detail for both the wet and dry laboratories. The methods and protocols presented in this volume will inspire the novice as well as the expert researcher to undertake new experimental endeavors in plant proteomics. Chapters 1 and 2 describe standard protocols and chapters 3 through 10 contain optimized protocols for protein extraction of tissues such as cereal seeds, xylem, phloem sap, of woody plants, and of cellular compartments, such as chloroplasts and mitochondria. These methods rely on improvements to solubilization of proteins, which is the emphasis of Chapters 11 and 12. Chapters 13 to 23 outline methods for protein identification, including 2DLC or LC of intact proteins, mass spectrometry (MALDI-TOF and MS/MS) with proteins separated by 2D gels, blue native ID gels, and Edman sequencing. Chapters 24 to 29 focus on the rapidly expanding area of analysis of post-translational modifications and protein-protein relationships. Attention is also given to data that proteomics produces, including management of that data, and to statistical tools used for effective analysis.
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