Mucins

This volume explores the latest advancements in mucin research. The chapters in this book are organized into 8 parts and cover a wide range of topics such as mucin extraction, isolation, physicochemical property analysis, and experimental methods. The chapters also discuss the origins of mucins in jellyfish, feces, saliva and salivary glands, bronchi, stomach, and cervical tract; organic synthesis of peptides glycosylated at specific sites; analysis of mucin gene expression and methylation-specific electrophoresis of genes; imaging of mucin networks by AFM; and experimental methods using supported molecular matrix electrophoresis. 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 tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Mucins: Methods and Protocols is a valuable resource for researchers who want to obtain in-depth insight on how to more thoroughly conduct mucin research in their laboratories.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Contenu
Preparation of Jellyfish Mucins.- Extraction of Mucin from Rodent Feces and Determination of O-Linked Oligosaccharide Chain Equivalent Derived from Fecal Mucin.- Preparation of Soluble Mucin Solutions from the Salivary Glands.- Isolation of Membrane Bound Mucins from Human Bronchial Epithelial Cells.- Extraction and Fractionation of Human Gastric Mucins from Gastric Juice.- Extraction of Mucins from the Mammalian Intestinal Tract.- Supported Molecular Matrix Electrophoresis.- Supported Immunohistochemical Staining of Mucins.- Succinylation-Alcian Blue Staining of Mucins on Polyvinylidene Diuoride Membrane.- Quantitation of Mucin by Densitometry of an Alcian Blue-Stained Membrane.- Quantitation of MUC5AC and MUC5B by Stable Isotope Labeling Mass Spectrometry.- Preparation of O-Glycans from Mucins using Hydrazine Treatment.- Eliminative Oximation of O-glycans from Mucins.- 9-Fluorenylmethyl Chloroformate Labeling forO-Glycan Analysis.- Liquid Chromatography and Capillary Electrophoresis Analysis of 2AA-Labeled O-Glycans.- Preparation of Mucin Glycopeptides by Organic Synthesis.- MALDI-TOF MS/MS Analysis of Permethylated O-Glycan Alditols Released from Mucins.- Structural Elucidation of Sialylated O-Glycan Alditols Obtained from Mucins by Mass Spectrometry.- Differential Glycoform Analysis of MUC1 Derived from Biological Specimens using an Antibody-Overlay Lectin Microarray.- ISOGlyP: O-Glycosylation Site Prediction using Peptide Sequences and GALNTs.- Assessment of Mucin-Associated Gene Expression Levels on the Ocular Surface.- Methylation Specific Electrophoresis.- Expression Analysis of Genes Corresponding to Mucins and their Glycans from Cervical Tissue using RNA Sequencing.- Recombinant Production of Glycoengineered Mucins in HEK293-F Cells.- Analysis of the Interaction between Mucin and Green Fluorescent Protein (GFP)-Tagged Galectin-2 using a 96-Well Plate.- Solution NMR Analysis of O-Glycopeptide-Antibody Interaction.- Cultivation of Microorganisms in Media Supplemented with Mucin Glycoproteins.- Bacterial Enzyme Assay for Mucin Glycan Degradation.- Measurement of Mucinase Activity.- Adhesion Inhibition Assay for Helicobacter pylori to Mucin by lactobacillus.- Imaging of Mucin Networks with Atomic Force Microscopy.- Molecular Dynamics Simulation and Docking of MUC1 O-Glycopeptide.- Fabrication and Characterization of Mucin Nanoparticles for Drug Delivery Applications.- Evaluation of Rheological Properties of Saliva by Determining the Spinnbarkeit.- Mechanical Characterization of Mucus on Intestinal Tissues by Atomic Force Microscopy.