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Min Li and a panel of hands-on experimentalists detail state-of-the-art molecular techniques for studying NMDA ligand-gated ion channels and developing assays for nontherapeutic lead selection. The topics range from cDNA cloning to in vitro and in vivo investigation of the channel complex in the mammalian brain. Additional topics include the biochemical analysis of the channel protein and the construction of various heterologous systems for both basic research and high throughput screens (HTS) for pharmaceutical chemicals. Although the focus is on NMDA receptors, the methods are applicable to other ligand-gated ion channels and with some modification may be extended to related membrane signaling receptors. NMDA Receptor Protocols offers today's scientists powerful methods for basic research on NMDA receptor structure and function, as well as enormous opportunities for clinical investigation toward the development of novel bioactive compounds.
Sommario
Isolation of Receptor Clones by Expression Screening in Xenopus Oocytes.- Expression of NMDA Receptor Channel Subunit Proteins Using Baculovirus and Herpesvirus Vectors.- Transient Expression of Functional NMDA Receptors in Mammalian Cells.- Stable Expression of Human NMDA Receptors in Cultured Mammalian Cells.- Expression of Extracellular N-Terminal Domain of NMDA Receptor in Mammalian Cells.- Immunocytochemistry of NMDA Receptors.- The Use of Ligand Binding in Assays of NMDA Receptor Function.- Calmodulin Modification of NMDA Receptors.- Detergent Solubilization and Immunoprecipitation of Native NMDA Receptors.- Redox Sensitivity of NMDA Receptors.- In Vitro Selection of Peptides Acting on NMDA Receptors.- NMDA Receptor Pharmacology and Analysis of Patch-Clamp Recordings.- Determination of Membrane Topology of Glutamate Receptors.- Engineering the NMDA Receptor Channel Lining.- High-Throughput Functional Detection of NMDA Receptor Activity.
Riassunto
Min Li and a panel of hands-on experimentalists detail state-of-the-art molecular techniques for studying NMDA ligand-gated ion channels and developing assays for nontherapeutic lead selection.
Relazione
"...In this volume of Methods in Molecular Biology great care has been taken to present the most modern approaches to solving the mysteries of the NMDA receptor...Each chapter includes detailed protocols and highly useful 'notes' that explain possible problems, sources of technical artifacts and how to overcome them....this book is informative and practical and would serve any channel or receptor researcher well." -Molecular Biotechnology
