Guide to Protein Purification

Guide to Protein Purification

2nd Edition - November 3, 2009

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  • Editors: Richard Burgess, Murray Deutscher
  • eBook ISBN: 9780080923178
  • Hardcover ISBN: 9780123745361

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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


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
    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

  • No. of pages: 900
  • Language: English
  • Copyright: © Academic Press 2009
  • Published: November 3, 2009
  • Imprint: Academic Press
  • eBook ISBN: 9780080923178
  • Hardcover ISBN: 9780123745361

About the Editors

Richard Burgess

Murray Deutscher

Affiliations and Expertise

University of Connecticut Health Center, Farmington, Connecticut, U.S.A.

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  • Ricard B. Fri Mar 01 2019

    Guide to Protein Purification

    Guide to Protein Purification