Guide to Protein Purification

1. Why Purify Enzymes?
Murray Deutscher and Arthur Kornberg

Section I: Developing Purification Procedures
2. Strategies and Considerations for Protein Purifications
Stuart Linn
3. Use of Bioinformatics in Planning a Protein Purification
Richard R. Burgess
4. Preparing A Purification Summary Table
Richard R. Burgess

Section II: General Methods for Handling Proteins and Enzymes
5. Setting Up A Laboratory
Murray P. Deutscher
6. Buffers: Principle and Practice
Vincent S. Stoll and John S. Blanchard
7. Measurement of Enzyme Activity
T. K. Harris and M. M. Keshwani
8. Quantitation of Protein
James E. Noble and Marc J. A. Bailey
9. Concentration of Proteins and Removal of Solutes
Jonathan K. Romero, David R. H. Evans and Matthew Westoby
10. Maintaining Protein Stability
Murray P. Deutscher

Section III: Recombinant Protein Expression and Purification

11. Selecting an Appropriate Method for Expressing a Recombinant Protein
William H. Brondyk
12. Bacterial Systems for Production of Heterologous Proteins
Frank R. Collart, Ashley M. Frank and Sarah Zerbs
13. Expression in the Yeast Pichia Pastoris
James M. Cregg, Ilya Tolstorukov, Anasua Kusari, Jay Sunga, Knut Madden and Thomas Chappell
14. Baculovirus-Insect Cell Expression Systems
Donald L. Jarvis
15. Recombinant Protein Production By Transient Gene Transfer Into Mammalian Cells
Sabine Geisse and Cornelia Fux
16. Tagging For Protein Expression
Arun Malhotra
17. Refolding of Solubilized Inclusion Body Proteins
Richard R. Burgess

Section IV: Preparation of Extracts and Subcellular Fractionation
18. Advances In Preparation of Biological Extracts for Protein Purification
Anthony C. Grabski
19. Isolation of Subcellular Organelles and Structures
Jörg Von Hagen and Uwe Michelsen

Section V: Purification Procedures: Bulk Methods
20. Protein Precipitation Techniques
Richard R. Burgess?
21. Affi-Gel Blue for Nucleic Acid Removal and Early Enrichment of Nucleotide Binding Protein
Murray P. Deutscher

Section VI: Purification Procedures: Chromatographic Methods
22. Ion Exchange Chromatography
Alois Jungbauer and Rainer Hahn
23. Gel Filtration
Earle Stellwagen
24. Protein Chromatography on Hydroxyapatite Columns
Larry J. Cummings, Mark A. Snyder and Kimberly Brisack
25. Theory and Use of Hydrophobic Interaction Chromatography in Protein Purification Applications
Justin T. Mccue

Section VII: Purification Procedures: Affininty Methods
26. Affinity Chromatography: General Methods
Marjeta Urh, Dan Simpson and Kate Zhao
27. Immobilized-Metal Affinity Chromatography (IMAC)
Frank Schäfer* Helena Block, Barbara Maertens, Anne Spriestersbach, Nicole Brinker, Jan Kubicek, Roland Fabis And Jörg Labahn
28. Identification, Production, and Use of Polyol-Responsive Monoclonal Antibodies for Immunoaffinity Chromatography
Nancy E. Thompson, Katherine M. Foley, Elizabeth S. Stalder, And Richard R. Burgess

Section VIII: Purification Procedures: Electrophoretic Methods
29. One-Dimensional Gel Electrophoresis
David E. Garfin
30. Protein Analysis Using High-Resolution Two-Dimensional Polyacrylamide Gel Electrophoresis and Isoelectric Focusing
Friedman
31. Protein Gel Staining Methods: An Introduction And Overview
Thomas H. Steinberg
32. Elution of Proteins From Gels
Richard R. Burgess
33. Immunodetection By Protein Blotting Performing and Optimizing Western Blots With An Emphasis on Chemiluminescent Detection
Alice Alegria-Schaffer, Krishna Vattem and Andrew Lodge

Section IX: Purification Procedures: Membrane Proteins And Glycoproteins
34. Detergents: An Overview
Dirk Linke
35. Purification of Membrane Proteins
Sue-Hwa Lin and Guido Guidotti
36. Purification of Recombinant G-Protein-Coupled Receptors
Reinhard Grisshammer
37. Cell-Free Translation Of Integral Membrane Proteins Into Unilamelar Liposomes
Michael A. Goren, Akira Nozawa, Shin-Ichi Makino, Russell L. Wrobel and Brian G. Fox

Section X: Characterization of Purified Proteins
38. Determination of Protein Purity
David G. Rhodes and Thomas M. Laue
39. Determination of Size, Molecular Weight, and Presence of Subunits
David G. Rhodes, Robert E. Bossio and Thomas M. Laue
40. Identification and Quantification of Protein Posttranslational Modifications
Andrew Link
41. Parallel Methods For Expression And Purification
Scott A. Lesley
42. Techniques To Isolate O2-Sensitive Proteins: [4Fe-4S]-FNR As An Example
Aixin Yan and Patricia J. Kiley
43. Rethinking Your Purification Procedure
Murray Deutscher
44. Important But Little Known (Or Forgotten) Artifacts in Protein Biochemistry
Richard R. Burgess