Neurotrophin Protocols
Neurotrophin Protocols
edited by Robert A. Rush.
- 1st edition 2001
- 1 recurso en línea (X, 274 páginas) 41 ilustraciones
- Methods in Molecular Biology 169 1940-6029 .
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.
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.
9781592590605
10.1385/1592590608 doi
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.
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.
9781592590605
10.1385/1592590608 doi