Doonan Protein Purification Protocols

Erscheinungsjahr 2008
ISBN: 978-1-59259-545-7
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 406 Seiten

Reihe: Springer Protocols

Hans Neurath has written that this is the second golden era of enzymology {Protein Science [1994], vol. 3, pp. 1734—1739); he could with justice have been more general and referred to the second golden age of protein chemistry. The last two decades have seen enormous advances in our understanding of the structures and functions of pro teins arising on the one hand from improvements and developments in analytical techniques {see the companion volume, Basic Protein and Peptide Protocols, in this series) and on the other hand from the tech nologies of molecular genetics. Far from turning the focus away from protein science, the ability to isolate, analyze, and express genes has increased interest in proteins as gene products. Hence, many laborato ries are now getting involved in protein isolation for the first time, either as an essential adjunct to their work in molecular genetics or because of a curiosity to know more about the products of the genes that they have been studying. Protein Purification Protocols is aimed mainly at these newcom ers to protein purification, but it is hoped that it will also be of value to established practitioners who may find here techniques that they have not tried, but which might well be most applicable in their work. With the exception mainly of the first and last chapters, the format of the contributions to the present book conform to the established format of the Methods in Molecular Biology series.

Zielgruppe

Research

Autoren/Hrsg.

Doonan, Shawn

Inhaltsverzeichnis

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.- Affinity Precipitation Methods.- Isoelectric Focusing.- Chromatofocusing.- Size-Exclusion Chromatography.- Fast Protein Liquid Chromatography (FPLC) Methods.- 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.- High-Performance Electrophoresis Chromatography.- Practical Column Chromatography.