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This volume provides a collection of protocols from leading researchers in the dynein field. The enclosed chapters guide readers through a broad range of powerful techniques that can be used to understand the complexities of dynein activity and regulation. These include how to perform live-cell microscopy on various fungi and metazoan cells, how to purify, assemble, and measure the force generation and motility characteristics of various dynein assemblies. 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.
Cutting-edge and thorough, Dynein: Methods and Protocols is a valuable resource for both novice and expert scientists in this developing field.
List of contents
Live-cell imaging of dynein-mediated cargo transport in Aspergillus nidulans.- Using budding yeast as a model to understand dynein-mediated cargo transport.- Employing live-cell imaging to study motor-mediated transport.- Assessment of dynein-mediated nuclear migration in the developing cortex by live-tissue microscopy.- Using optogenetics to spatially control cortical dynein activity in mitotic human cells.- Visualization of single dynein molecules in mammalian cells.- Single-molecule studies of motor adaptors using cell lysates.- Reconstitution of organelle transport along microtubules in vitro.- Reconstitution of dynein/dynactin transport using recombinant dynein.- Fast and easy transient mammalian cell expression and purification of cytoplasmic dynein.- High-resolution tracking of dynein-dynactin-BicD2 complexes.- In vivo trapping of latex bead phagosomes for quantitative force measurements.- Studying dynein mechanochemistry with an optical trap.- Measurements of theForce-Dependent Detachment Rates of Cytoplasmic Dynein from Microtubules.- NMR analysis of the interactions and conformational plasticity of dynein intermediate chain.- High-resolution Structural Analysis of Dyneins by Cryo-electron Microscopy.
