A Laboratory Guide to Glycoconjugate Analysis - Tapa blanda

Sinopsis

1 High-Performance Liquid Chromatography of Derivatized and Non-Derivatized Oligosaccharides: A Review.- 1.1 Introduction.- 1.2 Separation Techniques.- Size-exclusion chromatography.- Ion-exchange chromatography.- Hydrophilic-interaction chromatography.- Reversed-phase chromatography.- Mixed elution protocols.- 1.3 Detection Techniques.- Refractometer.- UV detector.- Fluorescence and photometric detection after derivatization.- Detection after Postcolumn Reaction.- Miscellaneous Detection Techniques.- References.- 2 Detection of the Presence of Carbohydrates in Glycoproteins.- 2.1 Introduction.- A general chemical based method for glycoprotein identification.- 2.2 Principle.- Substances interfering with detection.- 2.3 Glycoprotein Detection on Membranes or in Solution: Relative Merits of Protocols.- Membrane labelling.- Solution labelling.- Sensitivity of detection.- Initial experiments.- Interpretation of results and troubleshooting.- Additional analyses: Alternative detection techniques.- 2.4 Identification of Specific Monosaccharides or Glycans.- Lectins.- Monoclonal antibodies to carbohydrate determinants.- Glycosyl transferases.- Naturally occurring carbohydrate recognition proteins.- Glycosyl-phosphatidyl-inositol anchor detection.- 2.5 Detailed Analysis of Protein Glycosylation Release-Label-Profile.- Acknowlegements.- References.- 3 HPAE-PAD Monosaccharide and Oligosaccharide Analysis of Glycoproteins Electrotransferred onto Polyvinylidene Fluoride Membranes.- 3.1 Introduction.- 3.2 SDS-PAGE.- 3.3 Electroblotting and Staining.- 3.4 Monosaccharide Composition Analysis.- Chromatography, detection and quantification of monosaccharides.- 3.5 Oligosaccharide Mapping.- Acknowledgement.- References.- 4 Determination of the Disaccharide Composition of Glycosaminoglycans: Comparison of Chemical and Enzymatic Scission.- 4.1 Introduction.- 4.2 Technical Procedures.- Sample preparation.- 4.3 Troubleshooting.- Enzyme digests.- Chemical depolymerization.- HPLC analysis.- Acknowledgements.- References.- 5 Mass Spectrometric Analysis of Highly Acidic Polysaccharides.- 5.1 Introduction.- 5.2 Principles of Matrix-Assisted Laser Desorption.- 5.3 Technical Procedures.- Sample preparation.- 5.4 Troubleshooting.- References.- 6 Analysis of N-Glycans by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.- 6.1 Introduction.- 6.2 MALDI-MS: Principle of Operation.- Instrumentation.- Analytical protocols.- 6.3 Analysis of Glycoproteins.- Glycoprotein De-N-glycosylation with PNGase F.- MALDI-MS of glycoproteins.- 6.4 Analysis of Glycopeptides.- Structural analysis of N-glycans by MALDI-MS of glycopeptides in combination with exoglycosidase array sequencing.- MALDI-MS of glycopeptides.- 6.5 Analysis of Released Oligosaccharides.- Analysis of released N-glycans by MALDI-MS.- MALDI-MS of glycans.- 6.6 Analysing Carbohydrate Mass Information.- Quantitation.- Accessing the complex carbohydrate structure database.- Data for calculation of oligosaccharide masses.- References.- 7 Polyacrylamide Gel Electrophoresis of Fluorophore-Labelled Reducing Saccharides: A Review.- 7.1 Introduction.- 7.2 Principle.- 7.3 Preparation of Wheat Starch Digest Electrophoretic Standard.- Troubleshooting.- 7.4 Enzymatic Release of Asparagine-Linked Oligosaccharides from Glycoproteins Using PNGase F.- Troubleshooting.- 7.5 Release of Asparagine and Serine/Threonine-Linked Glycans from Glycoproteins Using Hydrazinolysis.- 7.6 Derivatization of the Oligosaccharides with ANTS.- 7.7 Derivatization of Oligosaccharides with AMAC.- 7.8 PAGE for the Separation of ANTS-Derivatized Oligosaccharides and for Acidic Oligosaccharides Derivatized with AMAC.- Troubleshooting.- 7.9 PAGE for the Separation of Neutral and Acidic AMAC-Derivatized Saccharides.- 7.10 Enzymatic Structural Analysis of N-Glycans.- 7.11 Viewing and Imaging the Electrofluorograms.- References.- 8 Carbohydrate Analysis with Capillary Electrophoresis.- 8.1 Introduction.- Instrumentation.- Capillaries.- 8.2 S

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