Cystic Fibrosis[: Methods and Protocols
William R. Skach
Springer Science & Business Media, 2002 - Medical - 652 pages
Since the cloning of the cystic fibrosis transmembrane conductance regulator (CFTR) nearly a decade ago, cystic fibrosis researchers, clinicians, and patients have come to rely increasingly on a diverse array of fundamental techniques to understand the molecular basis of this complex disease. Cystic Fibrosis Methods and Protocols consolidates a broad range of detailed and readily reproducible in vitro, cellular, and whole animal laboratory protocols into an indispensable resource. From electrophysiology and cell biology, to animal models and gene therapy, this comprehensive set of methods provides the step-by-step instructions needed for investigators to incorporate new approaches into their research programs. Specific protocols describe new techniques for diagnosis, in vitro methods for the expression and functional analysis of CFTR, novel biochemical and cellular systems to determine how mutations subvert CFTR function, and in vivo protocols to examine how CFTR dysfunction produces multisystem pathology in human and animal models. Comprehensive, multidisciplinary, and highly practical, Cystic Fibrosis Methods and Protocols makes accessible to today's cystic fibrosis investigator the powerful new scientific techniques required to investigate the basic science of the disease and to translate this into effective clinical solutions.
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acid activity aeruginosa agarose airway aliquots analysis antibody antimicrobial apical assay bacteria basolateral beads binding Biol blot buffer Cell Biology cell lines centrifuge centrosome chamber channel Chem chloride chloride channel complexes concentration containing culture cystic fibrosis cystic fibrosis transmembrane degradation detection dilution disease domain EDTA electrodes ENaC endocytosis epithelial cells exon fibrosis transmembrane conductance filter fluid fluorescence folding forskolin function gating gene genetic incubate infection inhibitors interactions lipid lung lysate medium method mg/mL mice microscope molecular monolayers mouse mutations NaCl Note oocytes patch clamp patients pellet peptide Physiol pipet plasma membrane plasmid plate protease proteasome protocol reaction reagent remove residues resuspended room temperature sample Sigma sodium solution sputum sterile studies Subheading supernatant surface syntaxin tissue transfection transmembrane conductance regulator transport Tris-HCl tube vector voltage wash Western blot wild-type xenograft yeast