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The past two decades have seen an expansion of interest in glycoproteins. From being a borderline area between carbohydrate and protein chemistry, it has become relevant to a wide range of biological phenomena. The aim of the book is to describe techniques which can be used to answer some of the basic questions about glycosylated proteins. Methods are discussed for isolation; compositional analysis; structure carbohydrate units; protein-carbohydrate linkages, keeping in mind the diverse nature of problems which readers may have to tackle.
Proteoglycans are some of the most elaborate macromolecules of mammalian and lower organisms. The covalent attachment of at least five types of glycosami- glycan side chains to more than forty individual protein cores makes these molecules quite complex and endows them with a multitude of biological functions. Proteoglycan Protocols offers a comprehensive and up-to-date collection of prepa- tive and analytical methods for the in-depth analysis of proteoglycans. Featuring st- by-step detailed protocols, this book will enable both novice and experienced researchers to isolate intact proteoglycans from tissues and cultured cells, to establish the composition of their carbohydrate moieties, to generate strategies for prokaryotic and eukaryotic expression, to utilize methods for the suppression of specific proteoglycan gene expression and for the detection of mutant cells and degradation products, and to study specific interactions between proteoglycans and extracellular matrix proteins as well as growth factors and their receptors. The readers will find concise, yet comprehensive techniques carefully drafted by leading experts in the field. Each chapter commences with a general Introduction, followed by a detailed Materials section, and an easy-to-follow Methods section. An asset of each chapter is the extensive notation that includes troubleshooting tips and practical considerations that are often lacking in formal methodology papers. The reader will find this section most valuable because it is clearly provided by experienced scientists who have first-hand knowledge of the techniques they outline. In addition, most of the chapters are well illustrated with examples of typical data generated with each method.
Sugar chains (glycans) are often attached to proteins and lipids and have multiple roles in the organization and function of all organisms. "Essentials of Glycobiology" describes their biogenesis and function and offers a useful gateway to the understanding of glycans.
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.
Due to the significant contributions of carbohydrates to the functional diversity of the cell, the challenging study of the glycome has expanded beyond the research of carbohydrate experts and into the wider scope of the life sciences. To aid all scientists now delving into this vital subject area, Glycomics: Methods and Protocols collects a compendium of detailed laboratory protocols reflecting the increasing availability of sample preparation, chromatographic, electrophoretic, mass spectrometric, and bioinformatic tools specifically designed for the analysis of glycosylation. Leading researchers in the field address subjects such as glycoprotein and proteoglycan analysis, glycosylation structure determination, as well as various approaches to investigate the interaction between glycans and a variety of carbohydrate-recognizing proteins in order to aid exploration into the functional significance of the oligosaccharides. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Glycomics: Methods and Protocols serves as a valuable guide for experimenters facing the challenges of glycan analysis in hope of providing further insights into the biology of cell-cell communication and interaction.
A guide to modern scanning electron microscopy instrumentation, methodology and techniques, highlighting novel applications to cell and molecular biology.