Guide to Protein Purification, Volume 463

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 Proteins

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

The 2e of this classic Guide to Protein Purification provides a complete update to existing methods in the field, reflecting the enormous advances made in the last two decades. In particular, proteomics, mass spectrometry, and DNA technology have revolutionized the field since the first edition’s publication but through all of the advancements, the purification of proteins is still an indispensable first step in understanding their function. This volume examines the most reliable, robust methods for researchers in biochemistry, molecular and cell biology, genetics, pharmacology and biotechnology and sets a standard for best practices in the field. It relates how these traditional and new cutting-edge methods connect to the explosive advancements in the field. This "Guide to" gives imminently practical advice to avoid costly mistakes in choosing a method and brings in perspective from the premier researchers while presents a comprehensive overview of the field today.

• Gathers top global authors from industry, medicine, and research fields across a wide variety of disciplines, including biochemistry, genetics, oncology, pharmacology, dermatology and immunology
• Assembles chapters on both common and less common relevant techniques
• Provides robust methods as well as an analysis of the advancements in the field that, for an individual investigator, can be a demanding and time-consuming process

Researchers, professors, and students of biochemistry, molecular biology and cell biology