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Full Description
This volume discusses detailed protocols for the analysis of glycosylation at the level of free glycans and glycopeptides. The book covers topics such as the importance of glycans in eukaryotic life (with a focus on mammals and particularly humans); recent technologies that allow the characterization of larger sets of samples; and method repeatability and robustness, as well as higher throughout with respect to sample preparation, measurement and data analysis. The chapters spans a wide range of techniques, including the analysis of fluorescently labeled glycans with HPLC, LC-MS analysis of glycopeptides both for glycosylation profiling and for in-depth tandem mass spectrometric analysis of protein glycosylation, and the analysis of glycans with fluorescent labeling, capillary electrophoresis in conjunction with laser induced fluorescence detection. The chapters also detail specific samples types including brain tissues, N-glycans from in-vitro cell cultures, milk oligosaccharides, invertebrate and protest N-glycans, as well as plant glycans. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction 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.
Practical and thorough, High-Throughout Glycomics and Glycoproteomics: Methods and Protocols is an essential reference for researchers planning to enter this rapidly evolving field.
Contents
Ubiquitous Importance of Protein Glycosylation.- Laboratory Experimental Design for a Glycomic Study.- High-Throughput Analysis of the IgG N-Glycome by UPLC-FLR.- High-Throughput Analysis of IgG Fc Glycopeptides by LC-MS.- Sialic Acid Derivatization for the Rapid Subclass and Sialic Acid Linkage-Specific MALDI-TOF-MS Analysis of IgG Fc-Glycopeptides.- The Use of Multiple Reaction Monitoring on QQQ-MS for the Analysis of Protein- and Site-Specific Glycosylation Patterns in Serum.- Analysis of Permethylated Glycan by Liquid Chromatography (LC) and Mass Spectrometry (MS).- High-Throughput Analysis of the Plasma N-Glycome by UHPLC.- Site-Specific N- and O-Glycopeptide Analysis Using an Integrated C18-PGC-LC-ESI-QTOF-MS/MS Approach.- Analysis of Milk Oligosaccharides by Mass Spectrometry.- N- and O-Glycomics from Minor Amounts of Formalin-Fixed, Paraffin-Embedded Tissue Samples.- Carbohydrate Microarray Technology Applied to High-Throughput Mapping of Plant Cell Wall Glycans Using Comprehensive Microarray Polymer Profiling (CoMPP).- Analysis of Invertebrate and Protist N-Glycans.- High-Throughput and High-Sensitivity Mass Spectrometry-Based N-Glycomics of Mammalian Cells.- Analysis of N-Glycosylation of Total Membrane Proteins.- HILIC-UPLC Analysis of Brain Tissue N-Glycans.- Automated Integration of a UPLC Glycomic Profile.- Databases and Associated Tools for Glycomics and Glycoproteomics.- High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation.
