1st Edition

Principles and Reactions of Protein Extraction, Purification, and Characterization

By Hafiz Ahmed Copyright 2004
    408 Pages 160 B/W Illustrations
    by CRC Press

    408 Pages
    by CRC Press

    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
    "Organized in a logical fashion…the most positive features in each chapter are the occasional placement of actual laboratory procedures (called "working procedures") and discussions on the advantages and disadvantages of each technique/method, often presented in tabular format. This comparison of methods and advice on when certain methods should be used is perhaps the most valuable feature of the book."
    - Biochemistry and Molecular Biology Education