Connexin Methods and Protocols (eBook, PDF)

Redaktion: Bruzzone, Roberto; Giaume, Christian

• Format: PDF

Produktbeschreibung

An essential element in direct cell-cell communication between virtually all cell types in vertebrates, connexins have been recently linked to an increasing number of human genetic diseases. In Connexin Methods and Protocols, well established investigators describe in detail their best techniques for studying the cellular and molecular biology of connexins, as well as their physiological properties. These forefront methods are specifically designed to investigate the life of connexins from biosynthesis to assembly into gap junction channels, and also include powerful assays to perform functional studies on these routes of cell-cell communication. To ensure experimental success, each time-tested technique contains detailed step-by-step instructions, as well as tips on how to avoid pitfalls. Also discussed are the many specific theoretical aspects of connexin biology and physiology. Cutting-edge and highly practical, Connexin Methods and Protocols offers laboratory researchers a comprehensive collection of proven methods for studying the role of connexins in intercellular communication and human genetic disorders, as well as finding promising new therapeutics for those disorders.

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Produktdetails

• Verlag: Humana Press
• Seitenzahl: 491
• Erscheinungstermin: 5. Februar 2008
• Englisch
• ISBN-13: 9781592590438
• Artikelnr.: 64612026

Inhaltsangabe

Tools to Study Connexins.- Investigation of Connexin Gene Expression Patterns by In Situ Hybridization Techniques.- Sodium Dodecyl Sulfate-Freeze-Fracture Immunolabeling of Gap Junctions.- Purification of Gap Junctions.- Culturing of Mammalian Cells Expressing Recombinant Connexins and Two-Dimensional Crystallization of the Isolated Gap Junctions.- Connexins/Connexons.- Biochemical Analysis of Connexon Assembly.- Expression and Imaging of Connexin-GFP Chimeras in Live Mammalian Cells.- Analysis of Connexin Expression in Brain Slices by Single-Cell Reverse Transcriptase Polymerase Chain Reaction.- Use of Retroviruses to Express Connexins.- Spatiotemporal Depletion of Connexins Using Antisense Oligonucleotides.- Transfection and Expression of Exogenous Connexins in Mammalian Cells.- Assays For Function.- Assaying the Molecular Permeability of Connexin Channels.- Applying the Xenopus Oocyte Expression System to the Analysis of Gap Junction Proteins.- Mutagenesis to Study Channel Structure.- Dual Patch Clamp.- Determining Ionic Permeabilities of Gap Junction Channels.- Fluorescence Recovery After Photobleaching.- Capture of Transjunctional Metabolites.- Physiology and Biology of Connexins.- The Study of Connexin Hemichannels (Connexons) in Xenopus Oocytes.- Exploring Hemichannel Permeability In Vitro.- Inducing De Novo Formation of Gap Juntion Channels.- Recording and Analysis of Putative Direct Electrical Interactions in the Mammalian Brain.- Intercellular Calcium Signaling and Flash Photolysis of Caged Compounds.- Biochemical Analysis of Connexin Phosphorylation.- How to Close a Gap Junction Channel.

Rezensionen

"This book will be invaluable to those in the connexin field and a fascinating read and useful text to those new to this area of research." -Molecular Biotechnology