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Main description:
Despite exciting advances in genome sequencing, isolating a protein from its expression system in its native form still presents a complex challenge. In High Throughput Protein Expression and Purification: Methods and Protocols, leading scientists detail the most successful protocols currently in use, including various high throughput cloning schemes, protein expression analysis, and production protocols. This volume describes the use of E. coli, insect, and mammalian cells, as well as cell-free systems for the production of a wide variety of proteins, including glycoproteins and membrane proteins, in order to best represent strategies that create and exploit common features to enable simplified cloning, stable expression, and purification of proteins. Written in the highly successful Methods in Molecular Biology (TM) series format, the chapters present brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section for tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and comprehensive, High Throughput Protein Expression and Purification: Methods and Protocols is an ideal reference for protein biochemists and all those who wish to apply these easy-to-use protocols to the many applicable fields.
Contents:
Part I: Reviews
1. High Throughput Protein Production (HTPP): A Review of Enabling Technologies to Expedite Protein Production
Jim Koehn and Ian Hunt
2. Designing Experiments for High-Throughput Protein Expression
Stephen P. Chambers and Susanne E. Swalley
Part II: Protocols for High-throughput Cloning
3. Gateway Cloning for Protein Expression
Dominic Esposito, Leslie A. Garvey, and Chacko S. Chakiath
4. Flexi Vector Cloning
Paul G. Blommel, Peter A. Martin, Kory D. Seder, Russell L. Wrobel, and Brian G. Fox
5. The Precise Engineering of Expression Vectors Using High Throughput In-Fusion (TM) PCR Cloning
Nick S. Berrow, David Alderton, and Raymond J Owens
6. The Polymerase Incomplete Primer Extension (PIPE) Method Applied to High-Throughput Cloning and Site-Directed Mutagenesis
Heath E. Klock and Scott A. Lesley
7. A Family of LIC Vectors for High-Throughput Cloning and Purification of Proteins
William H. Eschenfeldt, Lucy Stols, Cynthia Sanville Millard, Andrzej Joachimiak, and Mark I. Donnelly
8. 'System 48' High Throughput Cloning and Protein Expression Analysis
James M. Abdullah, Andrzej Joachimiak, and Frank R. Collart
Part III: Protocols for High-throughput Protein Expression and Purification
9. Automated 96-Well Purification of Hexahistidine-Tagged Recombinant Proteins on MagneHis Ni2+-Particles
Chiann-Tso Lin, Priscilla A. Moore, and Vladimir Kery
10. E.coli and Insect Cell Expression, Automated Purification and Quantitative Analysis
Stephen P. Chambers, John R. Fulghum, Douglas A. Austen, Fan Lu, and Susanne E. Swalley
11. Hexahistidine-Tagged Maltose-Binding Protein as a FusionPartner for the Production of Soluble Recombinant Proteins in Escherichia coli
Brian P. Austin, Sreedevi Nallamsetty, and David S. Waugh
12. PHB-Intein-Mediated Protein Purification Strategy
Alison R. Gillies, Mahmoud Reza Banki, and David W. Wood
13. High-Throughput Biotinylation of Proteins
Brian K. Kay, Sang Thai, and Veronica V. Volgina
14. High-Throughput Insect Cell Protein Expression Applications
Mirjam Buchs, Ernie Kim, Yann Pouliquen, Michael Sachs, Sabine Geisse, Marion Mahnke, and Ian Hunt
15. High-Throughput Protein Expression Using Cell-free System
Kalavathy Sitaraman and Deb K. Chatterjee
16. The Production of Glycoproteins by Transient Expression in Mammalian Cells
Joanne E. Nettleship, Nahid Rahman-Huq, and Raymond J. Owens
17. High-Throughput Expression and Detergent Screening of Integral Membrane Proteins
Said Eshaghi
18. Cell-Free Expression for Nanolipoprotein Particles (NLPs): Building a High-Throughput Membrane Protein Solubility Platform
Jenny A. Cappuccio, Angela K. Hinz, Edward A. Kuhn, Julia Fletcher, Erin S. Arroyo, Paul T. Henderson, Craig D. Blanchette, Vicki L. Walsowrth, Michele H. Corzett, Richard J. Law, Joseph B. Pesavento, Brent W. Segelke, Todd A. Sulchek, Brett A. Chromy, Federico Katzen, Todd Peterson, Graham Bench, Wieslaw Kudlicki, Paul D. Hoeprich Jr., and Matthew A. Coleman
19. Expression and Purification of Soluble His6-Tagged TEV Protease
Joseph E. Tropea, Scott Cherry, and David S. Waugh
20. High-Throughput Protein Concentration and Buffer Exchange: Comparison of Ultrafiltration and Ammonium Sulfate Precipitation
Priscilla A. Moore and Vladimir Kery
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 322
Weight: 829g
Availability: Available
Subcategories: Biochemistry
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