Protein Chromatography: Methods and Protocols
A prerequisite for elucidating the structure and function of any protein is the prior purification of that protein. This necessity has led to the development of many purification schemes and chromatographic methods for the isolation of native proteins from complex sources. In Protein Chromatography: Methods and Prools, leading researchers present clear prool-style chapters that are suitable for newcomers and experts alike. The book opens with vital topics in protein biochemistry, addressing such areas as protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the contrasting challenges that microfluidics and scale-up production pose to the investigator, and then it segues into key methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, amongst other means, as well as many variations on classic techniques such as ion-exchange and immunoaffinity chromatography. Written in the highly successful Methods in Molecular Biology™ series format, prools chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Protein Chromatography: Methods and Prools will greatly aid scientists in establishing these essential techniques in their own laboratories and furthering our understanding of the many imperative functions of proteins.
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Protein Chromatography: Methods and Protocols
A prerequisite for elucidating the structure and function of any protein is the prior purification of that protein. This necessity has led to the development of many purification schemes and chromatographic methods for the isolation of native proteins from complex sources. In Protein Chromatography: Methods and Prools, leading researchers present clear prool-style chapters that are suitable for newcomers and experts alike. The book opens with vital topics in protein biochemistry, addressing such areas as protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the contrasting challenges that microfluidics and scale-up production pose to the investigator, and then it segues into key methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, amongst other means, as well as many variations on classic techniques such as ion-exchange and immunoaffinity chromatography. Written in the highly successful Methods in Molecular Biology™ series format, prools chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Protein Chromatography: Methods and Prools will greatly aid scientists in establishing these essential techniques in their own laboratories and furthering our understanding of the many imperative functions of proteins.
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Protein Chromatography: Methods and Protocols

Protein Chromatography: Methods and Protocols

Protein Chromatography: Methods and Protocols

Protein Chromatography: Methods and Protocols

Hardcover(2011)

$169.99 
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Overview

A prerequisite for elucidating the structure and function of any protein is the prior purification of that protein. This necessity has led to the development of many purification schemes and chromatographic methods for the isolation of native proteins from complex sources. In Protein Chromatography: Methods and Prools, leading researchers present clear prool-style chapters that are suitable for newcomers and experts alike. The book opens with vital topics in protein biochemistry, addressing such areas as protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the contrasting challenges that microfluidics and scale-up production pose to the investigator, and then it segues into key methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, amongst other means, as well as many variations on classic techniques such as ion-exchange and immunoaffinity chromatography. Written in the highly successful Methods in Molecular Biology™ series format, prools chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Protein Chromatography: Methods and Prools will greatly aid scientists in establishing these essential techniques in their own laboratories and furthering our understanding of the many imperative functions of proteins.

Product Details

ISBN-13: 9781607619123
Publisher: Springer-Verlag New York, LLC
Publication date: 11/02/2010
Series: Methods in Molecular Biology , #681
Edition description: 2011
Pages: 527
Product dimensions: 7.30(w) x 10.10(h) x 1.50(d)

Table of Contents

Preface v

Contributors ix

Part I Overviews

1 A Digest of Protein Purification Dermot Walls Robert McGrath Sinéad T. Loughran 3

2 Gel-Filtration Chromatography Ciarán Ó'Fágáin Philip M. Cummins Brendan F. O'Connor 25

3 Immunoaffinity Chromatography Jenny Fitzgerald Paul Leonard Elaine Darcy Richard O'Kennedy 35

4 Avoiding Proteolysis During Protein Chromatography Barry J. Ryan 61

5 Scale-Up of Protein Purification: Downstream Processing Issues John Joseph Milne 73

6 Phage Display: A Powerful Technology for the Generation of High Specificity Affinity Reagents from Alternative Immune Sources William J. J. Finlay Laird Bloom Orla Cunningham 87

7 Engineering Protein Stability Ciarán Ó'Fágáin 103

8 Microfluidics in Protein Chromatography Frank A. Gomez 137

9 Tagging Recombinant Proteins to Enhance Solubility and Aid Purification Dermot Walls Sinéad T. Loughran 151

Part II Protocols

10 Storage and Lyophilisation of Pure Proteins Ciarán Ó'Fágáin 179

11 Differential Precipitation and Solubilization of Proteins Barry J. Ryan 203

12 Ion-Exchange Chromatography: Basic Principles and Application to the Partial Purification of Soluble Mammalian Prolyl Oligopeptidase Philip M. Cummins Oonagh Dowling Brendan F. O'Connor 215

13 Protein Quantitation and Analysis of Purity Eva M. Campion Sinéad T. Loughran Dermot Walls 229

14 Purification of Proteins Fused to Glutathione S-Transferase Sandra Harper David W. Speicher 259

15 Purification of Proteins Fused to Maltose-Binding Protein Mario Lebendiker Tsafi Danieli 281

16 Purification of Proteins from Baculovirus-Infected Insect Cells Luke O'Shaughnessy Sean Doyle 295

17 Purification of Poly-Histidine-Tagged Proteins Sinéad T. Loughran Dermot Walls 311

18 Immobilized Metal Affinity Chromatography/Reversed-Phase Enrichment of Phosphopeptides and Analysis by CID/ETD Tandem Mass Spectrometry Rosana Navajas Alberto Paradela Juan Pablo Albar 337

19 Tag Removal by Site-Specific Cleavage of Recombinant Fusion Proteins Adam Charlton Michael Zachariou 349

20 Purification of Antibodies Using Affinity Chromatography Elaine Darcy Paul Leonard Jenny Fitzgerald Martin Danaher Richard O'Kennedy 369

21 Optimized Generation of High-Affinity, High-Specificity Single-Chain Fv Antibodies from Multiantigen Immunized Chickens William J. J. Finlay Laird Bloom Orla Cunningham 383

22 Measuring Protein-Protein Interactions Using Biacore Paul Leonard Stephen Hearty Richard O'Kennedy 403

23 Ultra-Performance Liquid Chromatography-Mass Spectrometry of Proteins Timothy R. Croley 419

24 Hydrophobic Interaction Chromatography Philip M. Cummins Brendan F. O'Connor 431

25 Fast Protein Liquid Chromatography Ashkan Madadlou Siobhan O'Sullivan David Sheehan 439

26 Shotgun Proteomics: A Qualitative Approach Applying Isoelectric Focusing on Immobilized pH Gradient and LC-MS/MS Laurent Geiser Ali R. Vaezzadeh Jacques M. P. Deshusses Denis F. Hochstrasser 449

27 Shotgun Proteomics: A Relative Quantitative Approach Using Off-Gel Electrophoresis and LC-MS/MS Laurent Geiser Loïc Dayon Ali R. Vaezzadeh Denis F. Hochstrasser 459

28 Clinical Proteomics: Liquid Chromatography-Mass Spectrometry Purification Systems Michael Henry Paula Meleady 473

29 Strategies for the Purification of Membrane Proteins Sinead Marian Smith 485

30 A Multi-Step Chromatographic Strategy to Purify Three Fungal Endo-β-Glucanases Tracey McCarthy Maria G. Tuohy 497

Index 525

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