Title High resolution chromatography : a practical approach / edited by Paul Millner

Published Oxford ; New York : Oxford University Press, ©1999

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Description 1 online resource (xx, 311 pages) : illustrations Series Practical approach series ; 204 Practical approach series ; 204. Contents Intro; Contents; List of contributors; Abbreviations; Section A. Techniques and equipment; 1. Separation by charge, size, and hydrophobicity; 1. Introduction; 2. Choice of separation method; 3. Choice of hardware; 4. Packing columns; 5. Running the column; References; 2. Practice and benefits of microcolumn HPLC; 1. Introduction; 2. Practice of microcolumn HPLC; 3. Conclusion; References; 3. Detection devices; 1. Introduction; 2. The classification of detectors; 3. Detecting devices for high efficiency columns; 4. Light absorption detectors; 5. Fluorescence detectors; 6. Electronic detectors 7. Light scattering detectorsReferences; 4. Capillary electrophoresis of peptides and proteins; 1. Introduction; 2. The capillary; 3. Buffer; 4. Sample preparation; 5. Modes used in CE; 6. CE detection strategies; 7. Applications of CE to peptide and protein research; 8. CE of peptides and proteins: future prospects; References; Section B. Selected affinity approaches; 5. Lectins as affinity probes; 1. Properties and uses of lectins-a historical perspective; 2. The types of oligosaccharides found in glycoproteins; 3. Carbohydrate recognition and lectin specificity 4. Use of lectins: practical aspects5. Lectin affinity chromatography of oligosaccharides, glycosylasparagines, and glycopeptides; References; 6. Nucleotide- and dye-ligand chromatography; 1. Introduction; 2. Nucleotide-ligand chromatography; 3. Dye-ligand chromatography; References; 7. Synthetic peptides as affinity ligands; 1. Introduction; 2. Coupling peptides via amine groups directly to the matrix; 3. Coupling peptides via sulfydryl groups; 4. Coupling peptides and introducing spacer arms using carbodiimides; 5. Chromatography on peptide affinity gels; References 8. Drugs and inhibitors as affinity ligands1. Introduction; 2. Choice of ligand; 3. Choice of matrix; 4. Choice of spacer arm; 5. Coupling method; 6. Determination of ligand coupling efficiency; 7. Affinity chromatography; 8. Method of elution; 9. Affinity chromatography with biotinylated ligands; 10. Regeneration of the affinity matrix; Acknowledgements; References; 9. Immunoaffinity chromatography; 1. Introduction; 2. Antibody selection; 3. Applications of immunoaffinity separations; 4. Matrices; 5. Activation and immobilization; 6. Determining coupling efficiency 7. Equipment and operationReferences; 10. Strategies and methods for purification of Ca[sup(2+)]-binding proteins; 1. Introduction; 2. Purification of the annexin Ca[sup(2+)]-binding proteins; 3. The heparin column; 4. The Chelex-100 competitive Ca[sup(2+)]-binding assay; References; 11. Purification of DNA-binding proteins; 1. General procedure for DNA-binding proteins; 2. Preparation of nuclear extracts; 3. Heparin-Sepharose; 4. DNA affinity chromatography for purification of sequence specific DNA-binding proteins; References; Appendix; Index; A; B; C; D; E; F; G; H; I; L; M; N; O; P; Q; R Summary The molecular biology revolution has required the development of new chromatographic techniques and the optimization of original techniques to give reasonable quantities of protein at high resolutions. The aim of this volume is to provide the necessary information in most experimental situations to enable rapid and effective purification Bibliography Includes bibliographical references and index Notes English Subject Affinity chromatography. Biomolecules -- Purification Proteins -- Purification Chromatography, Affinity SCIENCE -- Chemistry -- Analytic. Affinity chromatography Biomolecules -- Purification Proteins -- Purification Affinitätschromatographie Chromatographie HPLC Kapillarelektrophorese Methode HPLC. Affiniteitschromatografie. Biomoleculen. Chromatographie d'affinité. Biomolécules -- Purification. Protéines -- Purification. Chromatographie en phase liquide à hautes performances. Form Electronic book Author Millner, Paul LC no. 98031925 ISBN 058548399X 9780585483993 9780191565663 0191565660 1280375477 9781280375477 9786610375479 661037547X

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