Potassium Channels: Methods and Protocols

ISBN-13: 9781617379406 / Angielski / Miękka / 2010 / 302 str.

Jonathan D. Lippiat

Opis

Due to the importance of potassium channels in cellular function, researchers have been challenged to develop tools and experimental procedures to probe the function and identify chemicals that modulate the behaviour of this super-family of ion channels. In Potassium Channels: Methods and Protocols, experts in the field present a range of experimental approaches that have been developed to investigate potassium channel structure, function, pharmacology, cell biology, gene expression, and their role in disease. Many of these techniques study potassium channels as cellular proteins as well as the resultant membrane biophysics. Written in the highly successful Methods in Molecular Biology series format, the chapters contain brief introductions to the subject, lists of the necessary materials and reagents, readily reproducible laboratory protocols, and a Notes section, which highlights tips on troubleshooting and avoiding known pitfalls. Thorough and cutting-edge, Potassium Channels: Methods and Protocols provides a wide range of approaches in order to aid current research projects and inspire new investigations."

Recenzje

From the reviews:

"This is a very informative book, covering a variety of methods available to study potassium channels ... . The protocols are clearly organized and usefully detailed. ... Illustrative figures and in some cases, photographs of equipment ... adding to the clarity of the methods sections. This book ... a valuable reference not only for those already in the potassium channel field but also for studies of other ion channel systems. ... each chapter allows bench scientists to undertake new, informed, experimental approaches in their research." (Samantha Miller, Microbiology Today, July, 2009)

"This book is dedicated to experimental methods, which allow us to investigate potassium channels from their genes through gene transcription to protein structure, function and regulation in health and disease. ... chapters in this book are written in form of practical laboratory protocols. Therefore the book will be primarily used by practicing scientists and by graduated and post-graduate students for their practical research. Certainly it will find its place in many laboratories investigating various features of potassium channels regulations." (L'ubica Lacinová, General Physiology and Biophysics, Vol. 29, April, 2010)

Spis treści

Part I: Methods for Studying Potassium Channel Transcription and mRNA Splicing Identifying Transcriptional Regulatory Regions Using Reporter Genes and DNA-Protein Interactions by Chromatin Immunoprecipitation Lezanne Ooi and Ian C. Wood Quantitative RT-PCR Methods for Investigation of Low Copy Potassium Channel Gene Expression in Native Murine Arteries Alex Cheong, Samuel J. Fountain, and David J. Beech Cloning of Potassium Channel Splice Variants from Tissues and Cells Lie Chen and Michael J. Shipston Part II: Methods for Studying the Cell Biology of Potassium Channel Proteins Chemiluminescence Assays to Investigate Membrane Expression and Clathrin Mediated Endocytosis of KATP Channels Andrew J. Smith and Asipu Sivaprasadarao Investigation of KATP Channel Endocytosis by Immunofluorescence Andrew J. Smith and Asipu Sivaprasadarao Investigation of KATP Channel Endocytosis and Cell Surface Density by Biotinylation and Western Blotting Andrew J. Smith and Asipu Sivaprasadarao Lipid Microdomains and K+ Channel Compartmentation: Detergent and Non-Detergent-Based Methods for the Isolation and Characterisation of Cholesterol-Enriched Lipid Rafts Laura J. Sampson and Caroline Dart Determination of Phosphoinositide Binding to K+ Channel Subunits Using a Protein Lipid Overlay Assay Alison M. Thomas and Andrew Tinker Protein Complex Analysis of Native Brain Potassium Channels by Proteomics Guillaume Sandoz and Florian Lesage Part III: Electrophysiological Techniques for the Study of Potassium Channel Function Xenopus Oocytes as a Heterologous Expression System for Studying Ion Channels with the Patch Clamp Technique Paolo Tammaro, Kenju Shimomura, and Peter Proks Whole Cell Recording Using the Perforated Patch Clamp Technique Jonathan D. Lippiat Recording the Activity of ATP-Sensitive K+ Channels in Open-Cell Cell-Attached Configuration Andrei I. Tarasov Planar Patch Clamp: Advances in Electrophysiology Andrea Brüggemann, Cecilia Farre, Claudia Haarmann, Ali Haythornthwaite, Mohamed Kreir, Sonja Stoelzle, Michael George, and Niels Fertig Analyzing Steroid Modulation of BKCa Channels Reconstituted into Planar Lipid Bilayers Heidi de Wet, Jonathan D. Lippiat, and Marcus Allen Part IV: Optical Techniques for the Study of Potassium Channel Function and Interactions Using Bioluminescence Resonance Energy Transfer (BRET) to Measure Ion Channel Assembly Gina M. Whitaker and Eric A. Accili The Use of FRET Microscopy to Elucidate Steady State Channel Conformational Rearrangements and G Protein Interaction with the GIRK Channels Adi Raveh, Inbal Riven, and Eitan Reuveny The Voltage-Clamp Fluorometry Technique Chris S. Ganhdi and Riccardo Olcese Part V: Methods for Studying Potassium Channelopathies and Pharmacological Modulators Identification of Mutations in the Kir6.2 Subunit of the KATP Channel Sarah E. Flanagan and Sian Ellard Modulation of Potassium Ion Channel Proteins Utilizing Antibodies Mark L Dallas, Susan A. Deuchars, and Jim Deuchars Fluorescence-Based Tl+-Influx Assays as a Novel Approach for Characterization of Small-Conductance Ca2+-Activated