Proteoglycans Methods and Protocols

This volume is a comprehensive and up-to-date collection of strategies, reproducible methods, and protocols for the in-depth analysis of Proteoglycans (PGs) and their glycan part, the GAGs. Chapters are divided into three parts detailing GAGs in biological specimens, protocols for the evaluation of the in vitro and in vivo effects of PGs/GAGs, and protocols for compounds related with the metabolic enzymes, epigenetic regulation, and PGs/GAGs-based inhibitors. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Cutting-edge and thorough, Proteoglycans: Methods and Protocols aims to provide information on the elucidated the structural and functional aspects of the complex matrix macromolecules such as the proteoglycans and glycosaminoglycans.

 

Detection of Glycosaminoglycans in Biological Specimens.- Immunochemical Detection and Glycosaminoglycan Disaccharide-Based Characterization Of Chondroitin Sulfate Proteoglycans.- Proteoglycans: A Tool for detecting hyaluronan by ELISA-like Methods.- Particle Exclusion Assay: A Tool for Measuring Hyaluronan Pericellular Matrix.- Urinary Glycosaminoglycans: Characterization and Quantification.- HPLC-MS characterization of tissue-derived heparan sulfate and chondroitin sulfate.- Evaluation of Glycosaminoglycans Biological Age in Cells and Tissues.- Studying the effects of Glycosaminoglycans in Cell Morphological Aspect With Scanning Electron Microscopy.- Evaluation of the In Vitro and In Vivo Effects of Biglycan in Innate Immunity.- Analysis of Extracellular Vesicle-associated Proteoglycans.- Analysis of Aggrecan Glycanation by Western Blot in Cell Culture.- Repulsive Scaling Replica Exchange Molecular Dynamics in ModelingProtein-Glycosaminoglycan Complexes.- In vitro investigation of hyaluronan/CD44 network.- Separation and Identification of Native Proteoglycans by Composite Agarose-Polyacrylamide Gel Electrophoresis and Immunoblotting.- Building, Visualizing, and Analyzing Glycosaminoglycan-Protein Interaction Networks.- An Enzymatic Activity Assay for Heparanase That Is Useful for Evaluating Clinically Relevant Inhibitors and Studying Kinetics.- A method for Urinary Trypsin Inhibitor (UTI) purification combining anion-exchange chromatography enrichment and preparative SDS-PAGE.- Enzymatic Sequencing of Heparin Oligosaccharides Using Exolyase.- Evaluating the Effects of microRNAs on Proteoglycans and Matrix Constituents’ Expression and Functional Properties.- Proteoglycan expression studied by microRNAs.- Modeling and simulations of Extra Cellular Glycoproteins.