Due to the importance of potassium channels in cellular function, researchershave been challenged to develop tools and 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. Contains detailed protocols that describe the use of routineand advanced electrophysiological techniques, such as whole cell patch clamp,perforated patch clamp, excised macropatch, open-cell cell-attached patch clamp, planar patch clamp, and artificial bilayer INDICE: From the contents Identifying Transcriptional Regulatory Regions Using Reporter Genes and DNA-Protein Interactions by Chromatin Immunoprecipitation.- Quantitative RT-PCR Methods for Investigation of Low Copy Potassium Channel Gene Expression in Native Murine Arteries.- Cloning of Potassium Channel Splice Variants from Tissues and Cells.- Chemiluminescence Assays to Investigate Membrane Expression and Clathrin Mediated Endocytosis of KATP Channels.- Investigation of KATP Channel Endocytosis by Immunofluorescence.- Investigation of KATP Channel Endocytosis and Cell Surface Density by Biotinylation and Western Blotting.- Lipid Microdomains and K+ Channel Compartmentation: Detergent and Non-Detergent-Based Methods for the Isolation and Characterisation of Cholesterol-Enriched Lipid Rafts.- Determination of Phosphoinositide Binding to K+ Channel Subunits Using a Protein Lipid Overlay Assay.- Protein Complex Analysisof Native Brain Potassium Channels by Proteomics.
- ISBN: 978-1-934115-65-7
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 295
- Fecha Publicación: 01/11/2008
- Nº Volúmenes: 1
- Idioma: Inglés