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Neurotrophin Protocols: Methods in Molecular Biology, cartea 169

Editat de Robert A. Rush
en Limba Engleză Paperback – 27 sep 2013
The past decade has seen an extraordinary growth in research interest in neurotrophic factors, and the study of the neurotrophin family has led this activity. Nevertheless, this area of research has often struggled as a result of techniques that were either inadequate or just emerging from other research fields and disciplines. Neurotrophin Protocols has brought together many leaders in the neurotrophin field who detail their special expertise in a wide variety of techniques. Though most procedures are valid across many diff- ent fields of research, some of those described here have been developed to address particular issues within the neurotrophic factor field. The protocols cover a broad range of biochemical, histological, and biological techniques that are often required by the modern laboratory. However, all have been written with sufficient detail to allow any laboratory to achieve proficiency without need of reference to other texts. Neurotrophin Protocols is divided into four sections dealing with p- tein, RNA, recombinant, and in vivo techniques. Protein techniques have in general been less successfully employed than those dealing with RNA or DNA. However, procedures that achieve localization and quantification of the neurotrophins are now being used more extensively. Their inclusion here should assist further studies at the protein level. Transgenic cell lines and animals are commonplace in the scientific research literature, but their inc- sion in several chapters in this book provide some novel uses that are not readily available elsewhere.
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Specificații

ISBN-13: 9781489939838
ISBN-10: 1489939830
Pagini: 288
Ilustrații: X, 274 p. 41 illus.
Dimensiuni: 155 x 235 x 15 mm
Greutate: 0.41 kg
Ediția:Softcover reprint of the original 1st ed. 2001
Editura: Humana Press Inc.
Colecția Humana
Seria Methods in Molecular Biology

Locul publicării:Totowa, NJ, United States

Public țintă

Research

Cuprins

Neurotrophin Quantification and Localization.- Localization of Neurotrophin Proteins Within the Central Nervous System by Immunohistochemistry.- Neurotrophin Immunohistochemistry in Peripheral Tissues.- Extraction and Quantification of the Neurotrophins.- Identifying Novel Proteins in Nervous Tissue Using Microsequencing Techniques.- RNA Quantification and Localization.- Quantification of mRNA Levels Using Northern Blotting.- Quantification of mRNA Levels Using Ribonuclease Protection Assay.- Quantification of Neurotrophin mRNA by RT-PCR.- Sensitive and Nonradioactive In Situ Detection of Neurotrophin mRNAs in the Nervous System.- Construction of Recombinant Proteins,and Transgenic Cells and Animals.- Expression of Neurotrophic Factors such as GDNF in the E. coli System.- Construction of Cells Expressing Neurotrophins.- Efficient Generation of Stable Pheochromocytoma (PC12) Cell Lines Using a Recombinant Retrovirus (LNC).- Construction and Analysis of Transgenic Animals Overexpressing Neurotrophins.- Construction of Transgenic Animals Overproducing Neurotrophins and Their Receptors.- In Vivo Analysis.- Tracing with Radiolabeled Neurotrophins.- Toxin-Induced Death of Neurotrophin-Sensitive Neurons.- Oligonucleotides as Inhibitors of Protein Synthesis.- The IODO-GEN Method for Labeling and the Use of Retrograde Axonal Transport to Assay Neurotrophins.- Stereological Estimation of Neuronal Number The Optical Disector/Cavalieri Combination.

Recenzii

"...provides a collection of chapters written by experts in developmental neuroscience, neuroanatomy, signaling mechanisms that regulate neuronal survival and plasticity, and neurodegenerative disorders....This book will be a valuable addition to libraries at Universities and various other institutes engaged in biomedical sciences, as well as to the personal collection of those scientists interested in growth factors....A strength of Neurotrophin Protocols is that it covers considerable ground with little repetition of background information and specific methods among chapters....we expect that this book will spend less time on the shelf and more time on lab benches and office desks. This is well done and timely contribution to the field of neurotrophic factor research."-Journal of Molecular Neuroscience

Textul de pe ultima copertă

Despite the extraordinary growth of research interest in neurotrophic factors, the techniques available have often been inadequate or just emergent from other disciplines. In Neurotrophin Protocols, established leaders in the neurotrophin field detail their special expertise in a wide variety of key protein, RNA, recombinant, and in vivo techniques. The protocols range from immunological analysis for the cellular localization and quantification of the neurotrophins, to genetic manipulation of cells and animals for the analysis of biological function, to quantitative analysis of the active neurotrophin genes. There are also radiotracing techniques for studying neurotrophin transport in both the retrograde and anterograde directions, procedures for using immunotoxins to study the effects of eliminating a single class of neurons, and the essential stereological method for estimation of neuronal numbers. Each method includes not only detailed step-by-step instructions, but also a list of necessary equipment and supplies, and valuable notes spelling out quick tips and tricks of the trade.
Timely and robust, Neurotrophin Protocols provides today's neuroscientists in both academia and industry with a comprehensive range of practical, readily reproducible methods for studying neurotrophins and the critically important effects they have on the nervous system.

Caracteristici

Includes supplementary material: sn.pub/extras