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Laboratory Methods in Vesicular and Vectorial Transport describes the procedures used to study the mechanisms of vesicular transport along the secretory and endocytic paths, including electron microscopy, autoradiography, and methods associated with cyto- and immunocytochemistry, genetics, and biochemistry. It investigates vectorial transport to the cisternal space of the rough endoplasmic reticulum (ER), as well as protein translocation across the ER, strategies for gaining access to the cytoplasm, cell-free analysis of vesicle fusion, the structure of glycoproteins, and the use of cell systems for analysis of vesicular traffic. Organized into seven parts encompassing 20 chapters, this volume begins with an overview of protein topology in the ER and the use of cross-linking methods to probe the molecular environment of translocating polypeptide chains. It then discusses the reconstitution of secretory protein translocation from detergent-solubilized rough microsomes; the use of anti-idiotype antibodies to characterize protein-protein interactions; the use of perforated cells to elucidate intracellular membrane transport; delivery of macromolecules into cells expressing a viral membrane fusion protein; and digitonin permeabilization procedures for studying endosome acidification and function. The reader is also introduced to reconstitution of intracellular vesicle fusion in a cell-free system after receptor-mediated endocytosis; immunoisolation using magnetic solid supports; endosome and lysosome purification by free-flow electrophoresis; remodeling of glycoprotein oligosaccharides after endocytosis; and replica plating of animal cells. This book will interest students, researchers, geneticists, biochemists, and cell biologists.
Methods in Plant Cell Biology provides in two volumes a comprehensive collection of analytical methods essential for researchers and students in the plant sciences. Individual chapters, written by experts in the field, provide an introductory overview, followed by a step-by-step technical description of the methods. This is accompanied by examples of typical results, illustrations, troubleshooting of potential pitfalls, sources of chemicals and equipment, and complete reference lists. Protocols are written to be easily comprehended by beginning research students, but these extensive volumes will also be a valuable addition to the libraries of expert researchers.Key Features* Written by experts, many of whom have developed the individual methods described* Contains most, if not all, the methods needed for modern research in plant cell biology* Up-to-date and comprehensive* Full references* Allows quick access to relevant journal articles and to the sources of chemicals required for the procedures* Selective concentration on higher plant methods allows for particular emphasis on those problems specific to plants
Methods in Plant Cell Biology provides in two volumes a comprehensive collection of analytical methods essential for researchers and students in the plant sciences. Individual chapters, written by experts in the field, provide an introductory overview, followed by a step-by-step technical description of the methods. Key Features* Written by experts, many of whom have developed the individual methods described* Contains most, if not all, the methods needed for modern research in plant cell biology* Up-to-date and comprehensive* Full references* Allows quick access to relevant journal articles and to the sources of chemicals required for the procedures* Selective concentration on higher plant methods allows for particular emphasis on those problems specific to plants
This new volume of Methods in Cell Biology looks at methods for analyzing of golgi complex function. Chapters cover such topics as in vitro reconstitution systems, fluorescence-based analysis of trafficking in mammalian cells and high content screening. With cutting-edge material, this comprehensive collection is intended to guide researchers for years to come. - Covers sections on model systems and functional studies, imaging-based approaches and emerging studies - Chapters are written by experts in the field - Cutting-edge material
Many cell biologists study the properties of cells grown outside of the animal, in the controlled environment of a culture dish. Many cell biologists would benefit from directing their attention to nerve cells, but are afraid to try growing nerve cells in culture because it has the reputation for being much more difficult and unreliable than growing other kinds of cells. This volume is written by cell biologists and neuroscientists who are experts at growing various kinds of neurons in culture and using them for diverse experimental applications. The aim of this volume is to demystify the techniques used to grow nerve cells and to show cell biologists that it is something that they can easily master. * Contains culture methodology for more than a dozen types of CNS and PNS neurons * Includes most recent and reliable techniques from expert practitioners for specific experimental applications * Addresses the latest strategies for transfecting neurons.
Cytometry is one of the most rapidly growing methodologies available for basic cell and molecular biology, cytogenetics, immunology, oncology, environmental sciences and also various fields of clinical medicine. This new edition, split into 2 Parts, is an almost completely new book, with nearly all of the chapters devoted to new topics. Like the previous volumes on cytometry published as part of the Methods in Cell Biology series, it provides a comprehensive description of particular cytometric methods and reviews their applications. Chapters present the theoretical foundations of the described methods, their applicability in experimental laboratory and clinical settings, and describes common traps and pitfalls such as problems with data interpretation, comparison with alternative assays, and choosing the optimal assay. - Comprehensive presentation of cytometric methods covering theoretical applications, applicability, potential pitfalls, and comparisions to alternative assays - Discusses many new assays developed since the previous edition - Presents recent developments in cytometric intrumentation/technology
Driven in part by the development of genomics, proteomics, and bioinformatics as new disciplines, there has been a tremendous resurgence of interest in physical methods to investigate macromolecular structure and function in the context of living cells. This volume in Methods in Cell Biology is devoted to biophysical techniques in vivo and their applications to cellular biology. Biophysical Tools for Biologists covers methods-oriented chapters on fundamental as well as cutting-edge techniques in molecular and cellular biophysics. This book is directed toward the broad audience of cell biologists, biophysicists, pharmacologists, and molecular biologists who employ classical and modern biophysical technologies or wish to expand their expertise to include such approaches. It will also interest the biomedical and biotechnology communities for biophysical characterization of drug formulations prior to FDA approval. - Describes techniques in the context of important biological problems - Delineates critical steps and potential pitfalls for each method
Driven in part by the development of genomics, proteomics, and bioinformatics as new disciplines, there has been a tremendous resurgence of interest in physical methods to investigate macromolecular structure and function in the context of living cells. This volume in Methods in Cell Biology is devoted to biophysical techniques in vitro and their applications to cellular biology. Biophysical Tools for Biologists covers methods-oriented chapters on fundamental as well as cutting-edge techniques in molecular and cellular biophysics. This book is directed toward the broad audience of cell biologists, biophysicists, pharmacologists, and molecular biologists who employ classical and modern biophysical technologies or wish to expand their expertise to include such approaches. It will also interest the biomedical and biotechnology communities for biophysical characterization of drug formulations prior to FDA approval. - Describes techniques in the context of important biological problems - Delineates critical steps and potential pitfalls for each method - Includes full-color plates to illustrate techniques
This book provides a practical guide to experimental methods for studying the development invertebrate deuterostomes as animal model systems. The chapters provide detailed experimental protocols that cover a broad range of topics in modern experimental methods. Topics covered range from rearing embryos to the care of adult animals, while also presenting the basic experimental methods including light and electron microscopy, used to study gene expression, transgenics, reverse genetics, and genomic approaches.* Covers a wide range of methods, from classical embryology through modern genomics* Discusses animals related to vertebrates, providing a valuable evolutionary perspective* Includes a practical guide to the use of sea urchins in the teaching laboratory
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 volume (complemented by the forthcoming companion volume by Cassimeris and Tran) brings together current research and protocols in the field of microtoubules 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 both experimental procedures and new developments in the field of microtubule in vitro research - Combines classical approaches and modern technologies - Presents easy-to-use protocols and thorough background information, compiled by leaders in the field