Guide to Protein Purification (2nd Ed.)
Methods in Enzymology Series, Vol. 463

Coordinators: Burgess Richard R, Deutscher Murray P.

Language: Anglais

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900 p. · 15.2x22.9 cm · Hardback

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
  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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