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This book presents a methodological introduction on clathrin-mediated endocytosis. Chapters guide readers through a number of model systems, purifications, reconstitutions, analysis techniques and systemic alterations of membrane function, which has been pivotal in understanding the role of clathrin and is panoply of interactors. 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, Clathrin-Mediated Endoytsis: Methods and Protocols aims to ensure successful results in the further study of this vital field.
List of contents
Purification of Clathrin-coated Vesicles from Adult Rat Brain.- Preparation of Synaptosomes from Mammalian Brain by Subcellular Fractionation and Gradient Centrifugation.- Probing Endocytosis during the Cell Cycle with Minimal Experimental Perturbation.- Assaying the Contribution of Membrane Tension to Clathrin-mediated Endocytosis.- Identifying Small Molecule Inhibitors of the Clathrin Terminal Domain.- Acute Manipulations of Clathrin-mediated Endocytosis at Presynaptic Nerve Terminals.- Imaging "Hot-wired" Clathrin-mediated Endocytosis.- Real-time Endocytosis Measurements by Membrane Capacitance Recording at Central Nerve Terminals.- Assaying Mutants of Clathrin Mediated Endocytosis in the Fly Eye.- Reconstitution of Clathrin Coat Disassembly for Fluorescence Microscopy and Single-molecule Analysis.- Spatial and Temporal Aspects of Phosphoinositides in Endocytosis Studied in Isolated Plasma Membranes.- SMrT Assay for Real-time Visualization and Analysis of Clathrin Assembly Reactions.- Real-time Monitoring of Clathrin Assembly Kinetics in a Reconstituted System.- Stimulated Emission Depletion (STED) Imaging of Clathrin-mediated Endocytosis in Living Cells.- Measuring Clathrin-coated Vesicle Formation with Single-molecule Resolution.- Cryo-electron Tomography of the Mammalian Synapse.- Quantitative Analysis of Clathrin-mediated Endocytosis in Yeast by Live Cell Fluorescence Microscopy.- Using FM Dyes to Monitor Clathrin-mediated Endocytosis in Primary Neuronal Culture.
