© 2004 • 801 pp., illus., appendices, index
Paperback • ISBN  0-87969-696-6

This title also available in: Hardcover

This manual complements Simpson’s Proteins and Proteomics manual, with a comprehensive collection of methods for protein purification from a variety of source preparations. The chapters include detailed protocols, methods for optimizing the performance of experiments, discussion of potential pitfalls, and troubleshooting advice.

SECTION I—Preparing Proteins for Proteomic Analysis

Chapter 1: Role of Separation Science in Proteomics

Richard J. Simpson

Chapter 2: Strategies for Purifying Proteins

Richard J. Simpson

SECTION II—Chromatographic Methods for Preparing Proteins for Proteomics

Chapter 3: Introduction to Chromatographic Methods for Protein and Peptide Purification

Richard J. Simpson

Chapter 4: Multidimensional Chromatography of Intact Proteins

Alex Apffel

Chapter 5: High–throughput Screening of Soluble Recombinant Proteins

Ting–Fang Wang and Andrew H.–J. Wang

Chapter 6: Ion–exchange Chromatography

Bengt Westerlund

Chapter 7: Size–exclusion Chromatography

Helena Hedlund

Chapter 8: Role of Reversed–phase High–performance Liquid Chromatography in Protein Isolation and Purification

C. David Carr and Robert L. Moritz

Chapter 9: Hydrophobic Interaction Chromatography

Richard J. Simpson and Paul A. O’Farrell

Chapter 10: Affinity and Immunoaffinity Chromatography

Keith Brocklehurst, Albert J. Courey, Sheraz Gul, Sue–Hwa Lin, and Robert L. Moritz

Chapter 11: Dye–Ligand Affinity Chromatography in Proteomic Research

Nikos E. Labrou

Chapter 12: Immobilized Metal–ion Affinity Chromatography

Lars Haneskog

Chapter 13: Protein Chromatography on Hydroxyapatite

Richard J. Simpson

Chapter 14: Chromatofocusing

Jamil Mohammad

SECTION III—Electrophoretic Methods for Preparing Proteins for Proteomics

Chapter 15: Introduction to Strategies Based on Electrophoresis for Separating Proteins

Richard J. Simpson

Chapter 16: Separation of Proteins Using Two–dimensional Gel Electrophoresis

Angelika Görg, Oliver Drews, and Walter Weiss

Chapter 17: Sample Preparation for High–resolution Two–dimensional Electrophoresis by Isoelectric Fractionation in an MCE

Ben R. Herbert, Pier Giorgio Righetti, John McCarthy, Jasmine Grinyer, Annalisa Castagna, Matthew Laver, Matthew Durack, Gerard Rummery, Robecca Harcourt, and Keith L. Williams

Chapter 18: An Electrophoresis Method for Purifying Proteins: The Gradiflow BF400 Instrument

Vicki L. Locke and Theresa M. Thomas

Chapter 19: Protein Purification Using Free–flow Electrophoresis

Peter J.A. Weber, Gerhard Weber, and Christoph Eckerskorn

SECTION IV—Analysis of Purified Proteins Prior to Structural Proteomics

Chapter 20: Introduction to Methods for Characterizing Purified Proteins

Richard J. Simpson

Chapter 21: Characterization of Proteins by Mass Spectrometry

Gavin E. Reid

Chapter 22: Analytical Ultracentrifugation: Protein Size, Conformation, and Interactions

Christopher A. MacRaild and Geoffrey J. Howlett

Chapter 23: Measuring the Conformational Stability of a Protein

Gerald R. Grimsley, Beatrice M.P. Huyghues–Despointes, C. Nick Pace and J. Martin Scholtz

Chapter 24: Combining Surface Plasmon Resonance with Mass Spectrometry: Identifying Binding Partners and Characterizing Interactions

Jos Buijs and Tohru Natsume

Chapter 25: Analysis of Carbohydrate from Glycoproteins

David Oxley, Graeme Currie, and Antony Bacic

APPENDICES

A1: An Overview of Protein Structure

Richard J. Simpson

A2: Measuring the Concentration of Proteins

Richard J. Simpson

A3: Concentrating Solutions of Protein

Richard J. Simpson

A4: Stabilizing Proteins for Storage

Richard J. Simpson

A5: One–dimensional Polyacrylamide Gel Electrophoresis

A6: Troubleshooting Guides

       HPLC

       2D Gels

A6: Cautions

Index