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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Total Protein Extraction with TCA-Acetone Valérie Méchin, Catherine Damerval and Michel Zivy Phenol Extraction of Proteins for Proteomic Studies of Recalcitrant Plant Tissues Mireille Faurobert, Esther Pelpoir and Jamila Chaïb Protein Extraction from Cereal Seeds Gérard Branlard and Emmanuelle Bancel Protein Extraction from Xylem and Phloem Sap Julia Kehr and Martijn Rep Protein Extraction from Woody Plants Christophe Plomion and Céline Lalanne Isolation of Chloroplast Proteins from Arabidopsis thaliana for Proteome Analysis Klaas J. van Wijk, Jean-Benoit Peltier and Lisa Giacomelli Isolation and Sub-Fractionation of Plant Mitochondria for Proteomic Analysis Holger Eubel, Joshua L. Heazlewood and A. Harvey Millar Extraction of Nuclear Proteins from Root Meristematic Cells Fernando González-Camacho and Francisco Javier Medina Isolation of Nuclear Proteins Setsuko Komatsu Isolation of Cell Wall Proteins from Medicago sativa Stems Bonnie S. Watson and Lloyd W. Sumner Plant Plasma Membrane Protein Extraction and Solubilization for Proteomic Analysis Véronique Santoni Detergents and Chaotropes for Protein Solubilization before Two-Dimensional Electrophoresis Thierry Rabilloud, Sylvie Luche, Véronique Santoni and Mireille Chevallet Two-Dimensional Electrophoresis for Plant Proteomics Walter Weiss and Angelika Görg Visible and Fluorescent Staining of Two-Dimensional Gels François Chevalier, Valérie Rofidal and Michel Rossignol Two-Dimensional Differential In-Gel Electrophoresis (DIGE) of Leaf and Roots of Lycopersicon esculentum Matthew Keeler, Jessica Letarte, Emily Hattrup, Fatimah Hickman and Paul A. Haynes Quantitative Analysis of 2D Gels Michel Zivy Multivariate Data Analysis of Proteome Data KåreEngkilde, Susanne Jacobsen and Ib Søndergaard Edman Sequencing of Plant Proteins from 2D Gels Setsuko Komatsu Peptide Mass Fingerprinting, Identification of Proteins by Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF) Nicolas Sommerer, Delphine Centeno and Michel Rossignol Protein Identification using Nano Liquid Chromatography: Tandem Mass Spectrometry Luc Negroni Two-Dimensional Nanoflow Liquid Chromatography: Tandem Mass Spectrometry of Proteins Extracted from Rice Leaves and Roots Linda Breci and Paul A. Haynes Separation, Identification and Profiling of Membrane Proteins by GFC/ IEC/SDS-PAGE and MALDI TOF MS Wojciech Szponarski, Frédéric Delom, Nicolas Sommerer, Michel Rossignol and Rémy Gibrat The PROTICdb Database for 2-DE Proteomics Olivier Langella, Michel Zivy and Johann Joets Identification of Phosphorylated Proteins Maria V. Turkina and Alexander V. Vener Plant Proteomics and Glycosylation Anne-Catherine Fitchette, Olivia Tran Dinh, Loïc Faye and Muriel Bardor Blue-Native Gel Electrophoresis for the Characterization of Protein Complexes in Plants Jesco Heinemeyer, Dagmar Lewejohann and Hans-Peter Braun Electroelution of Intact Proteins from SDS-PAGE Gels and Their Subsequent MALDI-TOF MS Analysis Zhentian Lei, Ajith Anand, Kirankumar S. Mysore and Lloyd W. Sumner Generation of Plant Protein Microarrays and Investigation of Antigen-Antibody Interactions Birgit Kersten and Tanja Feilner Phosphorylation Studies Using Plant Protein Microarrays Tanja Feilner and Birgit Kersten
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