Principles and Reactions of Protein Extraction, Purification, and Characterization By Hafiz Ahmed, Hafiz Ahmed PhD

Published September 14, 2004 by CRC Press

408 Pages 160 B/W Illustrations

Book Description

Principles and Reactions of Protein Extraction, Purification, and Characterization provides the mechanisms and experimental procedures for classic to cutting-edge techniques used in protein extraction, purification, and characterization.

The author presents the principles and reactions behind each procedure and uses tables to compare the different methods. The book also discusses the development of antibodies and immunochemical techniques as tools for characterizing proteins and modified proteins such as glycoproteins. Helpful illustrations, diagrams, and tables effectively transform theoretical concepts into practical knowledge. Along with methodical working procedures for most techniques, the book also offers useful advice on which technique to use and when to apply a particular method.

Presenting the advantages and disadvantages of the various protein techniques, Principles and Reactions of Protein Extraction, Purification, and Characterization enables students and researchers to master the mechanisms behind the protocols and choose the best method for their purposes.

EXTRACTION OF PROTEIN
Preparation of Buffers for Protein Extraction
Use of Protease Inhibitors in Protein Extraction
Use of Detergents in Protein Extraction
Chemical Lysis for Protein Extraction
Mechanical Lysis for Protein Extraction
Preparation of Extract from Prokaryotes
Extraction of Recombinant Protein from Bacteria
Preparation of Extracts from Yeast
Preparation of Extracts from Eukaryotes
Preparation of Extracts from Plants
Preparation of Membrane Extracts

ESTIMATION OF PROTEIN
Ultraviolet Absorption Methods
Colorimetric Methods
Fluorescent Methods

ELECTROPHORETIC ANALYSES OF PROTEIN
Driving Force of Electrophoresis
Polyacrylamide Gel Electrophoresis (PAGE)
Isoelectric Focusing (IEF)
Two-Dimensional (2-D) Gel Electrophoresis
Western Blotting
Capillary Electrophoresis

PURIFICATION OF PROTEIN
General Consideration and Purification Strategy
Non-Chromatographic Purification of Proteins
Chromatographic Purification of Proteins

ANTIBODIES: STRUCTURES, INTERACTIONS AND PRODUCTION
Immune System and Antibody Response
Structure of Antibodies
Antigen-Antibody Interactions
Production oof Antibodies
Development of Monoclonal Antibodies
Purification of Antibodies

ANTIBODY LABELING, ANTIBODY DETECTION AND IMMUNOCHEMICAL TECHNIQUES
Methods of Labeling Antibodies
Detection of Antibody Binding
Immunochemical Techniques: a Wide Range of Sensitive Assays for Detecting and Quantitating (Semi) Antigen-Antibody Reactions
Immunoprecipitation
Immunohistochemistry
Flow Cytometry

PURIFICATION OF GLYCOPROTEINS AND ANALYSES OF THEIR OLIGOSACCHARIDES
Diagrams and Stereochemistry of Monosaccharides and Oligosaccharides
Purification of Glycoproteins
Release of Oligosaccharides From Glycoproteins
Preparation of Glycopeptides by Proteolytic Cleavage
Estimation of Carbohydrates
Chromatographic Separation and Detection of Sugars
Electrophoretic Separation
Glycan Detection/Differentiation by Lectins
Analyses of Carbohydrate Linkage