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This second edition details techniques for the study of cargo trafficking through endosomes. New and updated chapters guide readers through methods and protocols on the structural aspects of plant endosomes, combined biochemical, omics, imaging approaches to study the dynamics and contents of endosomal compartments. Additional chapters are dedicated to the analysis of lipids on endosomes and the identification and analysis of lipid binding proteins and lipid-binding domains relevant for the study of plant endosomes. Written in the highly successful Methods in Molecular Biology series format, chapters include 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 cutting-edge, Plant Endosomes: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.
List of contents
Analysis of Endocytosis and Intracellular Trafficking of Boric Acid/Borate Transport Proteins in Arabidopsis.- Analysis of Membrane Proteins Transport from Endosomal Compartments to Vacuoles.- Analysis of Endoplasmic Reticulum-Endosomes Association Using Live-Cell Imaging In Plant Cells.- Degradation of Abscisic Acid Receptors through the Endosomal Pathway.- Biochemical and Imaging Analysis of ALIX Function in the Endosomal Trafficking of Arabidopsis Protein Cargoes.- Correlative Light and Electron Microscopy Imaging of the Plant trans-Golgi Network.- Imaging Plant Cells by High-pressure Freezing and Serial Block-Face Scanning Electron Microscopy.- Functional Analysis of Plant FYVE Domain Proteins in Endosomal Trafficking.- Assessing Extrinsic Membrane Protein Dependency to PI4P using a Plasma Membrane to Endosome Re-localisation Transient Assay in Nicotiana benthamiana.- Subcellular Localization of PI3P in Arabidopsis.- Immuno-purification of Intact Endosomal Compartments for Lipid Analyses in Arabidopsis.- Cell-free Protein Translation System For Expression Of Lipid-binding Proteins Tagged with Small Epitopes and their use in Protein-lipid Overlay Assays.- Isolation and Glycomic Analysis of Trans-Golgi Network Vesicles in Plants.- Detection of Phosphorylation on Immunoprecipitates from Total Protein Extracts of Arabidopsis thaliana Seedlings.- Purification and Interaction Analysis of a Plant-specific RAB5 Effector by in vitro pull-down Assay.
