Protein Purification Protocols

The first edition of Protein Purification Protocols (1996), edited by Professor Shawn Doonan, rapidly became very successful. Professor Doonan achieved his aims of p- ducing a list of protocols that were invaluable to newcomers in protein purification and of significant benefit to established practitioners. Each chapter was written by an ex- rienced expert in the field. In the intervening time, a number of advances have w- ranted a second edition. However, in attempting to encompass the recent developments in several areas, the intention has been to expand on the original format, retaining the concepts that made the initial edition so successful. This is reflected in the structure of this second edition. I am indebted to Professor Doonan for his involvement in this new edition and the continuity that this brings. Each chapter that appeared in the original volume has been reviewed and updated to reflect advances and bring the topic into the 21st century. In many cases, this reflects new applications or new matrices available from vendors. Many of these have increased the performance and/or scope of the given method. Several new chapters have been introduced, including chapters on all the currently used protein fractionation and ch- matographic techniques. They introduce the theory and background for each method, providing lists of the equipment and reagents required for their successful execution, as well as a detailed description of how each is performed.

Includes supplementary material: sn.pub/extras

Klappentext
In this new edition of the very successful Protein Purification Protocols (1996), Paul Cutler completely updates the existing protocols to reflect recent advances and adds an enormous new array of proteomic techniques for protein isolation and analysis. These cutting-edge techniques include not only two-dimensional gel electrophoresis for analysis and characterization, but also analytical chromatography for multidimensional separations of proteins and peptides, and mass spectrometry for isolating proteins. With the many recent advances in technology, simple spectrometric detection is no longer the only option for separating proteins, and the authors treat in full detail all the newer methods for these separations.
Comprehensive and highly practical, Protein Purification Protocols, Second Edition, brings together all the key methodologies that both novice and experienced investigators need to carry out successful experimental work on proteins and their functions today.

Inhalt
General Strategies.- Preparation of Extracts From Animal Tissues.- Protein Extraction From Plant Tissues.- Extraction of Recombinant Protein From Bacteria.- Protein Extraction From Fungi.- Subcellular Fractionation of Animal Tissues.- Subcellular Fractionation of Plant Tissues.- The Extraction of Enzymes From Plant Tissues Rich in Phenolic Compounds.- Avoidance of Proteolysis in Extracts.- Concentration of Extracts.- Making and Changing Buffers.- Purification and Concentration by Ultrafiltration.- Bulk Purification by Fractional precipitation.- Ion-Exchange Chromatography.- Hydrophobic Interaction Chromatography.- Affinity Chromatography.- Dye-Ligand Affinity Chromatography.- Lectin Affinity Chromatography.- Immunoaffinity Chromatography.- Immobilized Metal-Ion Affinity Chromatography.- Chromatography on Hydroxyapatite.- Thiophilic Affinity Chromatography and Related Methods.- Affinity Precipitation Methods.- Isoelectric Focusing.- Chromatofocusing.- Size-Exclusion Chromatography.- Fast Protein Liquid Chromatography.- Reversed-Phase Chromatography of Proteins.- Extraction of Membrane Proteins.- Removal of Detergent From Protein Fractions.- Purification of Membrane Proteins.- Lyophilization of Proteins.- Storage of Pure Proteins.- Electroelution of Proteins From Polyacrylamide Gels.- Electroblotting of Proteins From Polyacrylamide Gels.- Two-Dimensional Polyarcylamide Gel Electrophoresis for Proteome Analyses.- Microscale Solution Isoelectrofocusing.- Practical Column Chromatography.- Detection Methods.- Peptide Proteomics.- Multidimensional Liquid Chromatography of Proteins.- Mass Spectrometry.- Purification of Therapeutic Proteins.- Purification Process Scale-Up.