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Ion Channels (2nd Ed., 2nd ed. 2013) Methods and Protocols Methods in Molecular Biology Series, Vol. 998

Langue : Anglais

Coordonnateur : Gamper Nikita

Couverture de l’ouvrage Ion Channels

Every cell in our bodies contains a great variety and number of permeability pathways for various organic and inorganic ions, water, metabolites, nutrients, and signalling molecules. Maintenance and precise control of gating within these pathways are fundamental principles of life as these underlie basic cellular functions such as communication, contractility, and metabolism.  In Ion Channels: Methods and Protocols, Second Edition, experts in the field contribute chapters that focus on the strategies, approaches, methods, and protocols for studying a large family of proteins that form ionic channels in the plasma membrane and intracellular membranes of cells. Using practical examples from the cutting-edge current research, this volume will take a look back at the major methods and approaches that aided the progress toward the current understanding of ion channel function, structural design, and biological roles. The volume also aims to look forward and identify approaches that will lead us to future discoveries. Written in the highly successful Methods in Molecular Biology? series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Ion Channels: Methods and Protocols, Second Edition will greatly assist researchers searching for specific methodology in studying ion channels on the path toward a greater understanding of these key biological features.

Part I: Cloning and Heterologous Expression of Ion Channel Genes


1. Approaches to Cloning of Pain-Related Ion Channel Genes

            Armen N. Akopian


2. Mammalian Expression Systems and Transfection Techniques

            Daunia Laurenti and Lezanne Ooi


3. Use of Escherichia coli for the Production and Purification of Membrane Proteins

            Vincent G.L. Postis, Andrea E. Rawlings, Amelia Lesiuk, and Stephen A. Baldwin


4. Transient Overexpression of Genes in Neurons Using Nucleofection

            Hannah M. Kirton, Louisa Pettinger, and Nikita Gamper


5. Viral Gene Delivery: Optimized Protocol for Production of High Titre Lentiviral Vectors

            James Hewinson, Julian F.R. Paton, and Sergey Kasparov


Part II: Electrophysiological Methods to Study Ion Channel Functions


6. Two-Electrode Voltage Clamp

            Bingcai Guan, Xingjuan Chen, and Hailin Zhang


7. Conventional Micropipette-Based Patch Clamp Techniques

            Jonathan D. Lippiat and David C. Wrighton


8. Recording of Ion Channel Activity in Planar Lipid Bilayers Experiments

            Eleonora Zakharian


9. Recording Macroscopic Currents in Large Patches from Xenopus Oocytes

            Tibor Rohacs


10. Combined Single-Channel and Macroscopic Recording Techniques to Analyze Gating Mechanisms of the Large Conductance Ca2+ and Voltage Activated (BK) Potassium Channel

            Nguyen V. Nguyen, Aleksandra Gruslova, Wojciech A. Kosiba, and Bin Wang


11. Perforated Whole Cell Patch Clamp Recording

            John E. Linley


12. Piezo-Electrically Driven Mechanical Stimulation of Sensory Neurons

            Jizhe Hao, Jérôme Ruel, Bertrand Coste, Yann Roudaut, Marcel Crest, and Patrick Delmas


13. Automated Planar Patch-Clamp

            Carol J. Milligan and Clemens Möller


14. Recording Single-Channel Currents Using “Smart Patch-Clamp” Technique

            Anamika Bhargava and Julia Gorelik


Part III: Imaging and Fluorescence Methods to Study Ion Channels


15. Using Total Internal Reflection Fluorescence Microscopy to Observe Ion channel Trafficking and Assembly

            Sarah Schwarzer, Gregory I. Mashanov, Justin E. Molloy, and Andrew Tinker


16. Förster Resonance Energy Transfer-Based Imaging at the Cell Surface of Live Cells

            Sonya M. Bierbower and Mark S. Shapiro


17. The Use of Dansyl-Calmodulin to Study Interactions with Channels and Other Proteins

            Alessandro Alaimo, Covadonga Malo, Pilar Areso, Kerman Aloria, Oscar Millet, and Alvaro Villarroel


18. Imaging and Quantification of Recycled KATP Channels

            Christopher J. Cockcroft


Part IV: Biochemical and Structure-Functional Approaches in Ion Channel Studies


19. Generation of Antibodies that Are Externally-Acting Isoform-Specific Inhibitors of Ion Channels

            Jacqueline Naylor and David J. Beech


20. Site-Directed Mutagenesis to Study the Structure-Function Relationships of Ion Channels

            Wei Yang and Lin-Hua Jiang


21. Cysteine-Based Cross-Linking Approach to Study Inter-Domain Interactions in Ion Channel Functions

            Lin-Hua Jiang


22. Analysis of Ca2+-Binding Sites in the MthK RCK Domain by X-Ray Crystallography

            Frank J. Smith and Brad S. Rothberg


23. Isotope Labeling Strategies for Analysis of an Ion Channel Cytoplasmic Domain by NMR Spectroscopy

            Karin Abarca-Heidemann, Elke Duchardt-Ferner, Jens Woehnert, and Brad S. Rothberg


Part V: Studying Ion Channels in Native Tissues


24. Recording Dendritic Ion Channel Properties and Function from Cortical Neurons

            Mala M. Shah


25. M Current Recording from Acute DRG Slices

            Kirstin E. Rose, Sylvain Gigout, and Nikita Gamper


26. Studying Ion Channels in Human Erythrocytes by Direct and Indirect Means

            Stephan M. Huber


27. Recording Ion Channels in Isolated, Split-Opened Tubules

            Elena Mironova, Vladislav Bugay, Oleh Pochynyuk, Alexander Staruschenko, and James D. Stockand


28. Single Channel Analysis of TRPC Channels in the Podocytes of Freshly Isolated Glomeruli

            Daria V. Ilatovskaya and Alexander Staruschenko


29. Ca2+ Imaging as a Tool to Assess TRP Channel Function in Murine Distal Nephrons

            Mykola Mamenko, Oleg Zaika, Roger G. O’Neil, and Oleh Pochynyuk


30. Patch Clamping Drosophila Sensory Neurons

            Volodymyr Kucher, Benjamin A. Eaton, James D. Stockand, and Nina Boiko


Part VI: Ion Channels as Research Tools


31. Production and Validation of Recombinant Adeno-Associated Virus for Channelrhodopsin Expression in Neurons

            John Y. Lin


32. Optical Control of Ligand-Gated Ion Channels

            Stephanie Szobota, Catherine McKenzie, and Harald Janovjak

Includes both well-established and cutting-edge methods and protocols that focus on ionic channel function 

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

Date de parution :

Ouvrage de 440 p.

17.8x25.4 cm

Disponible chez l'éditeur (délai d'approvisionnement : 15 jours).

152,96 €

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Date de parution :

Ouvrage de 440 p.

17.8x25.4 cm

Disponible chez l'éditeur (délai d'approvisionnement : 15 jours).

179,34 €

Ajouter au panier
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