Guide to Protein Purification: Volume 436

Guide to Protein Purification: Volume 436

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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.
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Product details

  • Hardback | 900 pages
  • 154.94 x 226.06 x 40.64mm | 1,315.41g
  • Academic Press Inc
  • San Diego, United States
  • English
  • 2nd edition
  • 0123745365
  • 9780123745361

Table of contents

Why Purify Enzymes?
Murray Deutscher and Arthur Kornberg

Section I: Developing Purification Procedures

Strategies and Considerations for Protein Purifications
Stuart Linn

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

Preparing A Purification Summary Table
Richard R. Burgess

Section II: General Methods for Handling Proteins and Enzymes

Setting Up A Laboratory
Murray P. Deutscher

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

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

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

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

Maintaining Protein Stability
Murray P. Deutscher

Section III: Recombinant Protein Expression and Purification

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

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

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

Baculovirus-Insect Cell Expression Systems
Donald L. Jarvis

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

Tagging For Protein Expression
Arun Malhotra

Refolding of Solubilized Inclusion Body Proteins
Richard R. Burgess

Section IV: Preparation of Extracts and Subcellular Fractionation

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

Isolation of Subcellular Organelles and Structures
Joerg Von Hagen and Uwe Michelsen

Section V: Purification Procedures: Bulk Methods

Protein Precipitation Techniques
Richard R. Burgess

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

Section VI: Purification Procedures: Chromatographic Methods

Ion Exchange Chromatography
Alois Jungbauer and Rainer Hahn

Gel Filtration
Earle Stellwagen

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

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

Section VII: Purification Procedures: Affininty Methods

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

Immobilized-Metal Affinity Chromatography (IMAC)
Frank Schafer* Helena Block, Barbara Maertens, Anne Spriestersbach, Nicole Brinker, Jan Kubicek, Roland Fabis And Joerg Labahn

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

One-Dimensional Gel Electrophoresis
David E. Garfin

Protein Analysis Using High-Resolution Two-Dimensional Polyacrylamide Gel Electrophoresis and Isoelectric Focusing

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

Elution of Proteins From Gels
Richard R. Burgess

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

Detergents: An Overview
Dirk Linke

Purification of Membrane Proteins
Sue-Hwa Lin and Guido Guidotti

Purification of Recombinant G-Protein-Coupled Receptors
Reinhard Grisshammer

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

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

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

Identification and Quantification of Protein Posttranslational Modifications
Andrew Link

Parallel Methods For Expression And Purification
Scott A. Lesley

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

Rethinking Your Purification Procedure
Murray Deutscher

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

Richard R. Burgess
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