Cystic Fibrosis Methods and Protocols

Genetics of Cystic Fibrosis.- CFTR Mutation Detection by Multiplex Heteroduplex (mHET) Analysis on MDE Gel.- cDNA Microarrays for Pharmacogenomic Analysis of Cystic Fibrosis.- Natural Animal Models of Human Genetic Diseases.- CFTR structure and Function:.- Electrophysiological Approach to Studying CFTR.- Quantitative Analysis of ATP-Dependent Gating of CFTR.- CFTR Regulation by Phosphorylation.- Transepithelial Measurements of Bicarbonate Secretion in Calu-3 Cells.- Transepithelial Impedance Analysis of Chloride Secretion.- Studies of the Molecular Basis for Cystic Fibrosis Using Purified Reconstituted CFTR Protein.- Probing CFTR Channel Structure and Function Using the Substituted-Cysteine-Accessibility Method.- Methods for the Study of Intermolecular and Intramolecular Interactions Regulating CFTR Function.- Fluorescent Indicator Methods to Assay Functional CFTR Expression in Cells.- Immunolocalization of CFTR in Intact Tissue and Cultured Cells.- Analysis of CFTR Trafficking and Polarization Using Green Fluorescent Protein and Confocal Microscopy.- CFTR Folding and Maturation in Cells.- Isolation of CFTR.- CFTR Expression and ER-Associated Degradation in Yeast.- Manipulating the Folding Pathway of ?F508 CFTR Using Chemical Chaperones.- CFTR Degradation and Aggregation.- In Vitro Reconstitution of CFTR Biogenesis and Degradation.- In Vitro CFTR Folding Assays.- Analysis of CFTR Endocytosis by Cell Surface Biotinylation.- CFTR Regulation of ENaC.- Yeast Two-Hybrid Identification and Analysis of Protein Interactions with CFTR.- Biochemical Assays for Studying Indirect Interactions Between CFTR and the Cytoskeleton.- CFTR-Associated ATP Transport and Release.- Pathophysiology of Cystic Fibrosis.- Inflammatory Mediators in CF Patients.- Bacterial Colonization andInfection in the CF Lung.- Antimicrobial Peptides and Proteins in the CF Airway.- Bacterial-Epithelial Interactions.- Thin-Film Measurements of Airway Surface Liquid Volume/Composition and Mucus Transport

"The coverage of methodologies is extremely broad and will be useful for those wanting to find out more about some of the less routine techniques used in the analysis of the CFTR protein/chloride channel. The style of protocol-based chapters enables in manuscripts due to considerations of space or potential repetition of previously published techniques. As such many of the chapters are valuable." -Molecular Biotechnology

"Undoubtedly this book will be regarded as essential reading for scientists in the field. It provides details of research methodology, which are rarely published in peer reviewed journals. It will also enable researchers to identify subtle differences in techniques in their own laboratories from those described in these chapters. As such, it will be a very important book for all those who conduct laboratory-based research into cystic fibrosis." -Journal of Paediatrics and Child Health