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Full Description
Epithelial mucins are large complex cell surface and secreted glycoproteins produced by mucosal epithelial cells. In, Mucins: Methods and Protocols expert researchers in the field detail many of the methods which are now commonly used to study Mucins. These include methods and techniques for the best approaches to analysing each specific area of mucin biochemistry, physiology and biophysics before providing individual detailed experimental protocols together with troubleshooting and interpretation tips. 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 key tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Mucins: Methods and Protocols is designed to be a useful resource for those entering the mucin field and to facilitate those already studying mucins to broaden their experimental approaches to understanding mucosal biology.
Contents
Mucin Methods: Genes Encoding Mucins And Their Genetic Variation With A Focus On Gel Forming Mucins.- Gel-forming and Cell-associated Mucins - preparation for Structural and Functional Studies.- Detecting, Visualising and Quantifying Mucins.- Mass Spectrometric Analysis of Mucin Core Proteins.- O-Glycoprotein Biosynthesis: Site Localization by Edman Degradation and Site Prediction Based on Random Peptide Substrates.- Analysis of Assembly of Secreted Mucins.- MUC1 Membrane Trafficking: Protocols for Assessing Biosynthetic Delivery, Endocytosis, Recycling and Release Through Exosomes.- Glycomic Work-flow for Analysis of Mucin O-linked Oligosaccharides.- Glycomic Work-flow for Analysis of Mucin O-linked Oligosaccharides.- O-Glycomics: Profiling and Structural Analysis of Mucin-type O-linked Glycans.- O-Glycoproteomics: Site-specific O-glycoprotein Analysis by CID/ETD Electrospray Ionization Tandem Mass Spectrometry and Top-Down Glycoprotein Sequencing by In-Source Decay MALDI Mass Spectrometry.- Analysing Mucin Degradation.- Assessment of Mucus Thickness and Production In Situ.- Preservation of Mucus in Histological Sections, Immunostaining of Mucins in Fixed Tissue and Localization of Bacteria with FISH.- In vitro Measurements of Mucus Secretion by Colon Explants.- Establishment of Respiratory Air-Liquid Interface Cultures and their Use in Studying Mucin Production, Secretion, and Function.- Studying Mucin Secretion from Human Bronchial Epithelial Cell Primary Cultures.- Assessment of Intracellular Mucin Content In Vivo.- Techniques for Assessment of Interactions of Mucins with Microbes and Parasites In Vitroand In Vivo.- Assessing Mucin Expression and Function in Human Ocular Surface Epithelia In Vivo and In Vitro.
