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Guide to Protein Purification

  • 2nd Edition, Volume 463 - November 3, 2009
  • Latest edition
  • Editors: Richard R Burgess, Murray P. Deutscher
  • Language: English

Guide to Protein Purification, Second Edition provides a complete update to existing methods in the field, reflecting the enormous advances made in the last two decades. In partic… Read more

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Description

Guide to Protein Purification, Second Edition 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.

Key features

  • 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

Readership

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

Table of contents

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

Product details

  • Edition: 2
  • Latest edition
  • Volume: 463
  • Published: November 17, 2009
  • Language: English

About the editor

MD

Murray P. Deutscher

Affiliations and expertise
University of Connecticut Health Center, Farmington, Connecticut, U.S.A.

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