Exosomes and Microvesicles Methods and Protocols

This volume covers methods for the analysis of extracellular vesicles (EV) that can be applied to isolated EVs from a wide variety of sources. This includes the use of electron microscopy, tunable resistance pulse sensing, and nanoparticle tracking analysis. The chapters in this book discuss EV cargoes containing proteins and genomic materials using a number of different approaches, and isolating EVs from platelets and neuronal cells and tissues. 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.

Practical and comprehensive, Exosomes and Microvesicles: Methods and Protocols is a valuable resource containing methodologies for anyone interested in researching EVs.

Methods to Analyze EVs.- Tunable Resistive Pulse Sensing for the Characterization of Extracellular Vesicles.- Immuno-Characterization of Exosomes using Nanoparticle Tracking Analysis.- Imaging and Quantification of Extracellular Vesicles by Transmission Electron Microscopy.- Quantitative Analysis of Exosomal miRNA via qPCR and Digital PCR.- Small RNA Library Construction for Exosomal RNA from Biological Samples for the Ion Torrent PGM™ System.- A Protocol for Isolation and Proteomic Characterization of Distinct Extracellular Vesicle Subtypes by Sequential Centrifugal Ultrafiltration.- Multiplexed Phenotyping of Extracellular Vesicles using Protein Microarray (EV Array).- Purification and Analysis of Exosomes Released by Mature Cortical Neurons following Synaptic Activation.- A Method for Isolation of Extracellular Vesicles and Characterization of Exosomes from Brain Extracellular Space.- Isolation of Exosomes and Microvesicles from Cell Culture Systems to Study Prion Transmission.- Isolation of Platelet-Derived Extracellular Vesicles.- Bioinformatics Tools for Extracellular Vesicles Research.- Preparation and Isolation of siRNA-Loaded Extracellular Vesicles.- Interaction of Extracellular Vesicles with Endothelial Cells under Physiological Flow Conditions.