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This book brings together the major techniques used in the isolation or enrichment of individual populations of organelles and other subcellular structures from plants with the goal that, by being able to isolate subcellular structures, the research and understanding of various facets of compartmentalized function in plant cells can be advanced. Written for the highly successful Methods in Molecular Biology series, expert contributors provide chapters that contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Isolation of Plant Organelles and Structures: Methods and Protocols will greatly aid those who regularly isolate subcellular components as well as those whose research has lead them to focus on a subcellular compartment or a particular process for the first time, thus producing the need to be able to isolate it or enrich it for study.
Includes cutting-edge methods and protocols for the study of plant organelles and structures Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts
Inhalt
The Isolation of Plant Organelles and Structures in the Post-Genomics Era.- Approaches to Characterize Organelle, Compartment, or Structure Purity.- Isolation of Nuclei and Nucleoli.- Isolation and Suborganellar Fractionation of Arabidopsis Chloroplasts.- Isolation of Chromoplasts and Sub-Organellar Compartments from Tomato and Bell Pepper Fruit.- Leucoplast Isolation and Subfractionation.- Isolation of Mitochondria, Their Sub-Organellar Compartments, and Membranes.- Isolation of Arabidopsis Leaf Peroxisomes and the Peroxisomal Membrane.- Isolation of Vacuoles and the Tonoplast.- Isolation of Endoplasmic Reticulum and Its Membrane.- Enrichment of Golgi Membranes from Triticum aestivum (Wheat) Seedlings.- Isolation of Autolysosomes from Tobacco BY-2 Cells.- Isolation of Protein Storage Vacuoles and Their Membranes.- Isolation of the Cell Wall.- Isolation of Plasmodesmata.- Isolation of Plasma Membrane and Plasma Membrane Microdomains.- Enrichment of the Plant Cytosolic Fraction.- Isolation of Apoplast.- Isolation of Cytosolic Ribosomes.- Isolation of Plastid Ribosomes.- Isolation of Mitochondrial Ribosomes.- Isolation of Microtubules and Microtubule-Associated Proteins.- Isolation of Actin and Actin Binding Proteins.- Purification of 26S Proteasomes and Their Sub-Complexes from Plants.