There continues to be intense interest in the microtubule cytoskeleton; the assembly, structure and regulation of microtubules; and the numerous motors and accessory proteins that control cell cycle, dynamics, organization and transport. The field continues to grow and explore new aspects of these issues driven immensely by developments in optical imaging and tracking techniques. This 2e brings together current research and protocols in the field of microtubules in vitro and will serve as a valuable tool for cell biologists, biophysicists and pharmacologists who study the microtubule cytoskeleton, as well as for researchers in the biomedical and biotechnology communities with interest in developing drugs that target microtubules, MAPS and motors. Chapters reflect experimental procedures and new developments in the field of microtubule in vitro researchCombines classical approaches and modern technologiesPresents easy-to-use protocols and thorough background information, compiled by leaders in the field INDICE: Isolation of Antarctic fish tubulin and MAPsMaking, isolating and characterizing tubulin alpha antibodiesTechniques for labeling tubulin and drugsSingle-particle 3D reconstruction methods for decorated and undecorated microtubulesA new microtubule gliding assay and analysis of microtubule persistence lengthCharacterization of gamma tubulin Biochemical and Structural Studies of the doublecortin family of MAPsDetection and quantification of microtubule detachment from centrosomes and spindle polesRegulation of tubulin expression by micro RNAs: Implications for Drug ResistanceA portable magnetic tweezers device to visualize three-dimensional deformation of reconstituted microtubule networksMitochondrial/microtubule interactions - mmbp1Going Solo: Measuring the motions of microtubules with an in vitro assay for TIRF microscopyAnalysis of Microtubules and Motors in Isolated Axoplasm from the Squid Giant AxonAnalysis of drug binding to microtubules and steady state microtubule growth/treadmilling by use of a sedimentation assay in vitroSpecific recombinant antibody technology for the study of microtubule dynamics and regulatory proteinsTubulin-specific Chaperones: Components of a Molecular Machine that Assembles the aB ?HeterodimerG-protein tubulin interactionsKatanin, a microtubule severing enzyme: expression, purification, and biophysical characterizationMeasuring tubulin polymerization in vitro - comparison of methods and possible artifactsImaging microtubules in living plant cellsHow to use which antibody known in the field, what to expect from this antibody and how semi-quantitative data can be reliably obtained with antibody stainingPreparation of bacterial tubulin BtubA/B and constructs bearing eukaryotic tubulin sequencesMicrotubule-associated proteins and tubulin interaction by isothermal titration calorimetryTBCCharacterization of the mechanism of action of the covalent-binding microtubule-stabilizing agent zampanolideStudying kinetochore-fibre ultrastructure using correlative light-electron microscopyFluorescence-based Assays for Microtubule StructureYeast Tubulin-structure-function analysisMicrotubule Binding SIm to simulate binding of proteins to polymers under different binding models
- ISBN: 978-0-12-407757-7
- Editorial: Academic Press
- Encuadernacion: Cartoné
- Páginas: 440
- Fecha Publicación: 09/10/2013
- Nº Volúmenes: 1
- Idioma: Inglés