A comprehensive collection of essential, time-tested recipes for successful protein fractionation and purification in any experimental circumstance. The protocols give step-by-step instructions on how to select a source for the protein of interest, how to obtain a usable initial extract, how to purify the protein from that extract using both chemical and molecular methods, and how to dry and store the purified protein. Protein Purification Protocols provides all that is needed to design and carry out a successful purification program. It helps both experienced and novice investigators to clarify and define their purification problems and then provides a comprehensive set of tools for a practical solution.
General Strategies Shawn Doonan. Preparation of Extracts from Animal Tissues Shawn Doonan. Protein Extraction from Plant Tissues Peter R. Shewry and Roger J. Fido. Extraction of Recombinant Protein from Bacteria D. Margaret Worrall. Protein Extraction from Fungi Paul Bridge. Subcellular Fractionation of Animal Tissues Norma M. Ryan. Subcellular Fractionation of Plant Tissues: Isolation of Chloroplasts and Mitochondria from Leaves Alyson K. Tobin. The Extraction of Enzymes from Plant Tissues Rich in Phenolic Compounds William S. Pierpoint. Avoidance of Proteolysis in Extracts Robert J. Beynon and Simon Oliver. Concentration of Extracts Shawn Doonan. Making and Changing Buffers Shawn Doonan. Purification and Concentration by Ultrafiltration Paul Schratter. Bulk Purification by Fractional Precipitation Shawn Doonan. Ion- Exchange Chromatography David Sheehan and Richard FitzGerald. Hydrophobic Interaction Chromatography Paul O'Farrell. Affinity Chromatography Paul Cutler. Dye-Ligand Affinity Chromatography D. Margaret Worrall. Lectin Affinity Chromatography Iris West and Owen Goldring. Immunoaffinity Chromatography George W. Jack and David J. Beer. Immobilized Metal Ion Affinity Chromatography Tai-Tung Yip and T. William Hutchens. Chromatography on Hydroxyapatite Shawn Doonan. Affinity Precipitation Methods Jane A. Irwin and Keith F. Tipton. Isoelectric Focusing Reiner Westermeier. Chromatofocusing Timothy J. Mantle and Patricia Noone. Size-Exclusion Chromatography Paul Cutler. Fast Protein Liquid Chromatography (FPLC) Methods David Sheehan. Reversed-Phase Chromatography of Proteins Bill Neville. Extraction of Membrane Proteins Kay Ohlendieck. Removal of Detergent from Protein Fractions Kay Ohlendieck. Purification of Membrane Proteins Kay Ohlendieck. Lyophilization of Proteins Ciar n a F g in. Storage of Pure Proteins Ciar n a F g in. Electroelution of Proteins from Polyacrylamide Gels Michael J. Dunn. Electroblotting of Proteins from Polyacrylamide Gels Michael J. Dunn. High-Performance Electrophoresis Chromatography Serge Desnoyers Sylvie Bourassa and Guy G. Poirier. Practical Column Chromatography Shawn Doonan. Index.
...includes some very useful contributions. - Angewandte Chemie This book is recommended for protein chemists, whether beginner or professional. There is something in this book for everyone. It is an excellent working book as well as an excellent reference. - INFORM-American Oil Chemists Society Doonan's handy compendium...holds its own against Methods in Enzymology [and] Current protocols in Protein Science... will be valuable to keep around the laboratory for most graduate students in biochemistry, and indeed for more senior practitioners of biotechnology as well. - Biopharm Overall, a well presented utilitarian bench manual and a worthwhile purchase for novice and established practitioner alike - Society for General Microbiology Quarterly