Purifying proteins for proteomics : a laboratory manual / edited by Richard J. Simpson.

Format:

Book

Contributor:

Simpson, Richard J.

Language:

English

Subjects (All):

Proteins--Purification--Laboratory manuals.

Proteins.

Proteomics--Laboratory manuals.

Proteomics.

Proteins--isolation & purification.

Proteins--Purification.

Medical Subjects:

Proteins--isolation & purification.

Proteomics.

Genre:

Laboratory manuals.

Physical Description:

xi, 801 pages : illustrations (some color) ; 29 cm

Place of Publication:

Cold Spring Harbor, N.Y. : Cold Spring Harbor Laboratory Press, [2004]

Summary:

The first step in a proteomic analysis is the separation of a complex mixture of macromolecules into an array of simpler fractions. Purifying Proteins for Proteomics: A Laboratory Manual provides extensive background information, theory, protocols, and troubleshooting tips that will enable researchers to design and complete successful protein purification protocols using standard and cutting-edge techniques.

Contents:

Purifying Proteins for Proteomics Companion Web Site xi

Section I Preparing Proteins for Proteomic Analysis

1 Role of Separation Science in Proteomics / Richard J. Simpson 1

2 Strategies for Purifying Proteins / Richard J. Simpson 17

Section II Chromatographic Methods for Preparing Proteins for Proteomics

3 Introduction to Chromatographic Methods for Protein and Peptide Purification / Richard J. Simpson 41

4 Multidimensional Chromatography of Intact Proteins / Alex Apffel 75

5 High-throughput Screening of Soluble Recombinant Proteins / Ting-Fang Wang, Andrew H.-J. Wang 101

6 Ion-exchange Chromatography / Bengt Westerlund 121

7 Size-exclusion Chromatography / Helena Hedlund 147

8 Role of Reversed-phase High-performance Liquid Chromatography in Protein Isolation and Purification / C. David Carr, Robert L. Moritz 179

9 Hydrophobic Interaction Chromatography / Richard J. Simpson, Paul A. O'Farrell 209

10 Affinity and Immunoaffinity Chromatography / Keith Brocklehurst, Albert J. Courey, Sheraz Gul, Sue-Hwa Lin, Robert L. Moritz 221

11 Dye-Ligand Affinity Chromatography in Proteomic Research / Nikos E. Labrou 275

12 Immobilized Metal-ion Affinity Chromatography / Lars Haneskog 301

13 Protein Chromatography on Hydroxyapatite / Richard J. Simpson 337

14 Chromatofocusing / Jamil Mohammad 355

Section III Electrophoretic Methods for Preparing Proteins for Proteomics

15 Introduction to Strategies Based on Electrophoresis for Separating Proteins / Richard J. Simpson 381

16 Separation of Proteins Using Two-dimensional Gel Electrophoresis / Angelika Gorg, Oliver Drews, Walter Weiss 391

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, Keith L. Williams 431

18 An Electrophoresis Method for Purifying Proteins: The Gradiflow BF400 Instrument / Vicki L. Locke, Theresa M. Thomas 443

19 Protein Purification Using Free-flow Electrophoresis / Peter J.A. Weber, Gerhard Weber, Christoph Eckerskorn 463

Section IV Analysis of Purified Proteins prior to Structural Proteomics

20 Introduction to Methods for Characterizing Purified Proteins / Richard J. Simpson 479

21 Characterization of Proteins by Mass Spectrometry / Gavin E. Reid 489

22 Analytical Ultracentrifugation: Protein Size, Conformation, and Interactions / Christopher A. MacRaild, Geoffrey J. Howlett 517

23 Measuring the Conformational Stability of a Protein / Gerald R. Grimsley, Beatrice M.P. Huyghues-Despointes, C. Nick Pace, J. Martin Scholtz 535

24 Combining Surface Plasmon Resonance with Mass Spectrometry: Identifying Binding Partners and Characterizing Interactions / Jos Buijs, Tohru Natsume 567

25 Analysis of Carbohydrate from Glycoproteins / David Oxley, Graeme Currie, Antony Bacic 579

1. An Overview of Protein Structure / Richard J. Simpson 637

2. Measuring the Concentration of Proteins / Richard J. Simpson 659

3. Concentrating Solutions of Protein / Richard J. Simpson 701

4. Stabilization of Proteins for Storage / Richard J. Simpson 733

5. One-dimensional Polyacrylamide Gel Electrophoresis 751

6. Troubleshooting Guides 765

HPLC

2D Gels

7. Cautions 777.

Notes:

Includes bibliographical references and index.

ISBN:

0879696958

0879696966

OCLC:

52942962