Guide to Protein Purification book cover

Guide to Protein Purification

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.

Audience

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

Included in series
Methods in Enzymology

Hardbound, 900 Pages

Published: November 2009

Imprint: Academic Press

ISBN: 978-0-12-374536-1

Contents

    1. Why Purify Enzymes?
    2. Murray Deutscher and Arthur Kornberg

      Section I: Developing Purification Procedures

    3. Strategies and Considerations for Protein Purifications
    4. Stuart Linn

    5. Use of Bioinformatics in Planning a Protein Purification
    6. Richard R. Burgess

    7. Preparing A Purification Summary Table
    8. Richard R. Burgess

      Section II: General Methods for Handling Proteins and Enzymes

    9. Setting Up A Laboratory
    10. Murray P. Deutscher

    11. Buffers: Principle and Practice
    12. Vincent S. Stoll and John S. Blanchard

    13. Measurement of Enzyme Activity
    14. T. K. Harris and M. M. Keshwani

    15. Quantitation of Protein
    16. James E. Noble and Marc J. A. Bailey

    17. Concentration of Proteins and Removal of Solutes
    18. Jonathan K. Romero, David R. H. Evans and Matthew Westoby

    19. Maintaining Protein Stability
    20. Murray P. Deutscher

      Section III: Recombinant Protein Expression and Purification

    21. Selecting an Appropriate Method for Expressing a Recombinant Protein
    22. William H. Brondyk

    23. Bacterial Systems for Production of Heterologous Proteins
    24. Frank R. Collart, Ashley M. Frank and Sarah Zerbs

    25. Expression in the Yeast Pichia Pastoris
    26. James M. Cregg, Ilya Tolstorukov, Anasua Kusari, Jay Sunga, Knut Madden and Thomas Chappell

    27. Baculovirus-Insect Cell Expression Systems
    28. Donald L. Jarvis

    29. Recombinant Protein Production By Transient Gene Transfer Into Mammalian Cells
    30. Sabine Geisse and Cornelia Fux

    31. Tagging For Protein Expression
    32. Arun Malhotra

    33. Refolding of Solubilized Inclusion Body Proteins
    34. Richard R. Burgess

      Section IV: Preparation of Extracts and Subcellular Fractionation

    35. Advances In Preparation of Biological Extracts for Protein Purification
    36. Anthony C. Grabski

    37. Isolation of Subcellular Organelles and Structures
    38. Jörg Von Hagen and Uwe Michelsen

      Section V: Purification Procedures: Bulk Methods

    39. Protein Precipitation Techniques
    40. Richard R. Burgess

    41. Affi-Gel Blue for Nucleic Acid Removal and Early Enrichment of Nucleotide Binding Proteins
    42. Murray P. Deutscher

      Section VI: Purification Procedures: Chromatographic Methods

    43. Ion Exchange Chromatography
    44. Alois Jungbauer and Rainer Hahn

    45. Gel Filtration
    46. Earle Stellwagen

    47. Protein Chromatography on Hydroxyapatite Columns
    48. Larry J. Cummings, Mark A. Snyder and Kimberly Brisack

    49. Theory and Use of Hydrophobic Interaction Chromatography in Protein Purification Applications
    50. Justin T. Mccue

      Section VII: Purification Procedures: Affininty Methods

    51. Affinity Chromatography: General Methods
    52. Marjeta Urh, Dan Simpson and Kate Zhao

    53. Immobilized-Metal Affinity Chromatography (IMAC)
    54. Frank Schäfer* Helena Block, Barbara Maertens, Anne Spriestersbach, Nicole Brinker, Jan Kubicek, Roland Fabis And Jörg Labahn

    55. Identification, Production, and Use of Polyol-Responsive Monoclonal Antibodies for Immunoaffinity Chromatography
    56. Nancy E. Thompson, Katherine M. Foley, Elizabeth S. Stalder, And Richard R. Burgess

      Section VIII: Purification Procedures: Electrophoretic Methods

    57. One-Dimensional Gel Electrophoresis
    58. David E. Garfin

    59. Protein Analysis Using High-Resolution Two-Dimensional Polyacrylamide Gel Electrophoresis and Isoelectric Focusing
    60. Friedman

    61. Protein Gel Staining Methods: An Introduction And Overview
    62. Thomas H. Steinberg

    63. Elution of Proteins From Gels
    64. Richard R. Burgess

    65. Immunodetection By Protein Blotting Performing and Optimizing Western Blots With An Emphasis on Chemiluminescent Detection
    66. Alice Alegria-Schaffer, Krishna Vattem and Andrew Lodge

      Section IX: Purification Procedures: Membrane Proteins And Glycoproteins

    67. Detergents: An Overview
    68. Dirk Linke

    69. Purification of Membrane Proteins
    70. Sue-Hwa Lin and Guido Guidotti

    71. Purification of Recombinant G-Protein-Coupled Receptors
    72. Reinhard Grisshammer

    73. Cell-Free Translation Of Integral Membrane Proteins Into Unilamelar Liposomes
    74. Michael A. Goren, Akira Nozawa, Shin-Ichi Makino, Russell L. Wrobel and Brian G. Fox

      Section X: Characterization of Purified Proteins

    75. Determination of Protein Purity
    76. David G. Rhodes and Thomas M. Laue

    77. Determination of Size, Molecular Weight, and Presence of Subunits
    78. David G. Rhodes, Robert E. Bossio and Thomas M. Laue

    79. Identification and Quantification of Protein Posttranslational Modifications
    80. Andrew Link

    81. Parallel Methods For Expression And Purification
    82. Scott A. Lesley

    83. Techniques To Isolate O2-Sensitive Proteins: [4Fe-4S]-FNR As An Example
    84. Aixin Yan and Patricia J. Kiley

    85. Rethinking Your Purification Procedure
    86. Murray Deutscher

    87. Important But Little Known (Or Forgotten) Artifacts in Protein Biochemistry

    Richard R. Burgess

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