Guide to Protein Purification, 2nd Edition

✍ Scribed by Richard R. Burgess and Murray P. Deutscher (Eds.)

• Publisher: Academic Press
• Year: 2009
• Tongue: English
• Leaves: 853
• Series: Methods in Enzymology 463
• Edition: 2
• Category: Library

✦ Synopsis

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

✦ Table of Contents

Content:
Series Editors
Pages i-iii

Copyright Page
Page iv

Contents
Pages v-xvii

Contributors to Volume 463
Pages xix-xxiv

Preface
Pages xxv-xxvi
Richard R. Burgess, Murray P. Deutscher

Volume in Series
Pages xxvii-liii

Preface to Chapter 1
Page liv
Murray P. Deutscher

Chapter 1 Why Purify Enzymes?
Pages 3-6
Arthur Kornberg

Chapter 2 Strategies and Considerations for Protein Purifications
Pages 9-19
Stuart Linn

Chapter 3 Use of Bioinformatics in Planning a Protein Purification
Pages 21-28
Richard R. Burgess

Chapter 4 Preparing a Purification Summary Table
Pages 29-34
Richard R. Burgess

Chapter 5 Setting Up a Laboratory
Pages 37-42
Murray P. Deutscher

Chapter 6 Buffers: Principles and Practice
Pages 43-56
Vincent S. Stoll, John S. Blanchard

Chapter 7 Measurement of Enzyme Activity
Pages 57-71
T.K. Harris, M.M. Keshwani

Chapter 8 Quantitation of Protein
Pages 73-95
James E. Noble, Marc J.A. Bailey

Chapter 9 Concentration of Proteins and Removal of Solutes
Pages 97-120
David R.H. Evans, Jonathan K. Romero, Matthew Westoby

Chapter 10 Maintaining Protein Stability
Pages 121-127
Murray P. Deutscher

Chapter 11 Selecting an Appropriate Method for Expressing a Recombinant Protein
Pages 131-147
William H. Brondyk

Chapter 12 Bacterial Systems for Production of Heterologous Proteins
Pages 149-168
Sarah Zerbs, Ashley M. Frank, Frank R. Collart

Chapter 13 Expression in the Yeast Pichia pastoris
Pages 169-189
James M. Cregg, Ilya Tolstorukov, Anasua Kusari, Jay Sunga, Knut Madden, Thomas Chappell

Chapter 14 Baculovirus–Insect Cell Expression Systems
Pages 191-222
Donald L. Jarvis

Chapter 15 Recombinant Protein Production by Transient Gene Transfer into Mammalian Cells
Pages 223-238
Sabine Geisse, Cornelia Fux

Chapter 16 Tagging for Protein Expression
Pages 239-258
Arun Malhotra

Chapter 17 Refolding Solubilized Inclusion Body Proteins
Pages 259-282
Richard R. Burgess

Chapter 18 Advances in Preparation of Biological Extracts for Protein Purification
Pages 285-303
Anthony C. Grabski

Chapter 19 Isolation of Subcellular Organelles and Structures
Pages 305-328
Uwe Michelsen, Jörg von Hagen

Chapter 20 Protein Precipitation Techniques
Pages 331-342
Richard R. Burgess

Chapter 21 Affi-Gel Blue for Nucleic Acid Removal and Early Enrichment of Nucleotide Binding Proteins
Pages 343-345
Murray P. Deutscher

Chapter 22 Ion-Exchange Chromatography
Pages 349-371
Alois Jungbauer, Rainer Hahn

Chapter 23 Gel Filtration
Pages 373-385
Earle Stellwagen

Chapter 24 Protein Chromatography on Hydroxyapatite Columns
Pages 387-404
Larry J. Cummings, Mark A. Snyder, Kimberly Brisack

Chapter 25 Theory and Use of Hydrophobic Interaction Chromatography in Protein Purification Applications
Pages 405-414
Justin T. McCue

Chapter 26 Affinity Chromatography: General Methods
Pages 417-438
Marjeta Urh, Dan Simpson, Kate Zhao

Chapter 27 Immobilized-Metal Affinity Chromatography (IMAC): A Review
Pages 439-473
Helena Block, Barbara Maertens, Anne Spriestersbach, Nicole Brinker, Jan Kubicek, Roland Fabis, Jörg Labahn, Frank Schäfer

Chapter 28 Identification, Production, and Use of Polyol-Responsive Monoclonal Antibodies for Immunoaffinity Chromatography
Pages 475-494
Nancy E. Thompson, Katherine M. Foley, Elizabeth S. Stalder, Richard R. Burgess

Chapter 29 One-Dimensional Gel Electrophoresis
Pages 497-513
David E. Garfin

Chapter 30 Isoelectric Focusing and Two-Dimensional Gel Electrophoresis
Pages 515-540
David B. Friedman, Sjouke Hoving, Reiner Westermeier

Chapter 31 Protein Gel Staining Methods: An Introduction and Overview
Pages 541-563
Thomas H. Steinberg

Chapter 32 Elution of Proteins from Gels
Pages 565-572
Richard R. Burgess

Chapter 33 Performing and Optimizing Western Blots with an Emphasis on Chemiluminescent Detection
Pages 573-599
Alice Alegria-Schaffer, Andrew Lodge, Krishna Vattem

Chapter 34 Detergents: An Overview
Pages 603-617
Dirk Linke

Chapter 35 Purification of Membrane Proteins
Pages 619-629
Sue-Hwa Lin, Guido Guidotti

Chapter 36 Purification of Recombinant G-Protein-Coupled Receptors
Pages 631-645
Reinhard Grisshammer

Chapter 37 Cell-Free Translation of Integral Membrane Proteins into Unilamelar Liposomes
Pages 647-673
Michael A. Goren, Akira Nozawa, Shin-ichi Makino, Russell L. Wrobel, Brian G. Fox

Chapter 38 Determination of Protein Purity
Pages 677-689
David G. Rhodes, Thomas M. Laue

Chapter 39 Determination of Size, Molecular Weight, and Presence of Subunits
Pages 691-723
David G. Rhodes, Robert E. Bossio, Thomas M. Laue

Chapter 40 Identification and Quantification of Protein Posttranslational Modifications
Pages 725-763
Adam R. Farley, Andrew J. Link

Chapter 41 Parallel Methods for Expression and Purification
Pages 767-785
Scott A. Lesley

Chapter 42 Techniques to Isolate O₂-Sensitive Proteins: [4Fe–4S]-FNR as an Example
Pages 787-805
Aixin Yan, Patricia J. Kiley

Chapter 43 Rethinking Your Purification Procedure
Pages 809-811
Murray P. Deutscher

Chapter 44 Important but Little Known (or Forgotten) Artifacts in Protein Biochemistry
Pages 813-820
Richard R. Burgess

Author Index
Pages 821-833

Subject Index
Pages 835-851