Immunochemistry LabFax

Immunochemistry LabFax

1st Edition - March 7, 1994

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  • Editors: B. Hames, D. Rickwood
  • eBook ISBN: 9780080984438

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LABFAX volumes are purpose-designed data reference books for practicing scientists. Each book presents key information for a major subject in one place and so saves hours of searching. The authors and editors of each LABFAX volume have searched the original literature for the accurate data which they know the specialist needs.Immunochemistry Labfax is a detailed compendium of essential information - on plasma proteins, immunoglobulin properties and purification, antibody production, labeling and derivatization, plus data on techniques such as ELISA, blotting, and immunolocalization - needed constantly by researchers in any area of immunology and related disciplines.

Table of Contents

  • Contributors


    1. Plasma Proteins

    Plasma and Serum


    Properties of Commonly Used Anticoagulants (Table 1)


    Preparation of Serum from Plasma (Reversal of Anticoagulants) (Table 2)

    Plasma/Serum Proteins

    Human Blood Plasma Proteins in Binding and Transport of Ligands (Table 3)

    Human Blood Plasma Apolipoproteins (Table 4)

    Human Blood Plasma Proteins in Coagulation and Fibrinolysis (Table 5)

    Human Blood Plasma Proteinase Inhibitors (Table 6)

    Human Immunoglobulins (Table 7)

    Human Complement Components (Table 8)

    Factors Affecting the Concentration of Plasma Proteins

    The Effect of Age

    Protein, Albumin, γ-Globulin Plasma Concentrations (Table 9)

    Concentration of Immunoglobulin Isotypes in Plasma (Table 10)

    Concentration of IgG Subclasses in Plasma (Table 11)

    Concentration of C3, C4 and C5 in Plasma (Table 12)

    The Acute Phase Response

    Acute Phase Proteins (Table 13)

    Plasma Proteins in Other Body Fluids

    Concentrations of Immunoglobulins and Albumin in Body Fluids (Table 14)

    Antibodies Recognizing Plasma/Serum Proteins

    Absorption of Unwanted Specificities

    Anti-Whole Sera Antibodies

    Purified Antibodies

    IgG fractions

    Affinity Purified Antibodies

    Monoclonal Antibodies

    Storage of Commercial Antibodies


    2. Properties of Immunoglobulins


    Diversity of Immunoglobulins

    The General Structure of Immunoglobulins (Figure 1)

    General Properties of Human Immunoglobulins (Table 1)

    Polypeptide Composition of Human Immunoglobulins (Table 2)

    Distribution and Characteristics of Human Immunoglobulins (Table 3)

    Biological Properties of Human IgG Subclasses (Table 4)

    Reported Antibody Specificities of Human IgG Subclasses (Table 5)

    Biological Properties of Human IgA Subclasses (Table 6)

    Human Immunoglobulin Allotypes

    Specificity and Frequency of Human Immunoglobulin Allotypes (Table 7)

    Immunoglobulins from Other Species

    Properties of Mouse Immunoglobulins (Table 8)

    Properties of Rat Immunoglobulins (Table 9)

    Distribution of Light Chain Types Among Various Species (Table 10)

    Immunoglobulin Receptors (Fc Receptors)

    Molecular Properties of Human Leukocyte Fc Receptors (Table 11)

    Distribution and Regulation of Human Leukocyte Fc Receptor Expression (Table 12)

    General Characteristics of Other Fc Receptors (Table 13)


    3. Antibodies Recognizing Immunoglobulins

    The Specificity of Antibodies Recognizing Immunoglobulins of Other Species

    Anti-Immunoglobulin Whole Molecule Antibodies

    Anti-Immunoglobulin Heavy or Light Chain-Specific Antibodies

    Antibodies Against Immunoglobulin Subclasses

    Cross-Reactions of Anti-Immunoglobulin Reagents

    Conjugated and Derivatized Anti-Immunoglobulin Reagents

    Commercial Availability of Antibodies Recognizing Immunoglobulins of Different Species


    4. Producing Antibodies

    Polyclonal Antisera

    Preparation of Immunogen

    Haptenization of Peptides


    Species and Immunization Routes

    Immunization Protocols

    Bleeding Animals and Preparation and Storage of Sera

    Monoclonal Antibodies

    'Immune' Lymphocytes

    Cell fusion (Hybridoma Production) and Fusion Partners

    Cell Culture, Medium and Supplements


    Production of Monoclonal Antibodies

    Production of Human Monoclonal Antibodies by Lymphocyte Transformation

    rDNA-Produced Antibodies

    Spliced Immunoglobulin Molecules

    Recombinant DNA-Derived Immunoglobulin Fragments


    5. Purification and Fragmentation of Immunoglobulins

    General Properties of Antibodies


    Sources of Immunoglobulins

    Sources of Immunoglobulins (Table 2)

    Methods of Purification of IgG

    Precipitation with Sodium Sulfate or Ammonium Sulfate

    Precipitation with Sodium Sulfate or Ammonium Sulfate Followed by Ion Exchange Chromatography

    Precipitation with Caprylic Acid (Octanoic Acid)

    Precipitation with Caprylic Acid Followed by Ion Exchange Chromatography

    Chromatography on Immobilized Protein A or Protein G

    Purification of IgM

    Gel Filtration

    Purification of Serum IgA

    Precipitation with Ammonium Sulfate Followed by Ion Exchange Chromatography

    Chromatography on Immobilized Jacalin

    Other Techniques for Immunoglobulin or Antibody Purification

    Chromatography on Immobilized Anti-Immunoglobulin

    Chromatography on Immobilized Antigen

    Properties of Bacterial IgG-Binding Proteins: Protein A and protein G

    Properties of IgA-Binding Proteins

    Purified Immunoglobulins Available Commercially

    Preparation of Immunoglobulin Fragments


    6. Preparation and Use of Radiolabeled Antibodies and Antigens

    Labeling with Radioisotopes

    Labeling with Iodine-125

    Labeling with Other Isotopes

    Production of Radiolabeled Antibodies for Radioscintography

    Labeling with ß-emitter (Tritium)



    Assay Components

    Data Analysis

    Further Optimization

    Immunoradiometric Assay

    Assay Components

    Data Analysis

    Assay Optimization


    7. Enzyme-Conjugated Antibodies: Preparation and Use


    Enhanced Enzyme Immunodetection Systems

    Enzymes Commonly Used in Immunoconjugates

    Horseradish Peroxidase

    Alkaline Phosphatase


    Glucose Oxidase


    Methods for the Conjugation of Enzymes to Antibodies

    Glutaraldehyde-Mediated Conjugation

    Conjugation Mediated by Heterobifunctional Cross-Linking Reagents

    Periodate-Mediated Conjugation

    Properties of Enzyme Substrates Commonly Used in Immunoassay

    Substrates for Horseradish Peroxidase

    Substrates for Alkaline Phosphatase

    Substrates for ß-Galactosidase

    Substrates for Glucose Oxidase

    Substrates for Urease


    8. Derivatization of Antibodies and Antigens: Flourescent Labeling, Biotinylation, Immunogold Derivatives

    Fluorescently Labeled Antibodies

    Choice of Fluorochrome

    Conjugation of Antibodies

    Laser-Excitation of Fluorochromes

    Biotinylation of Proteins


    Properties of Biotin

    Properties of Avidin

    Properties of Streptavidin

    Colloidal Gold Derivatives

    Properties of Colloidal Gold Conjugates


    9. Common Immunological Techniques: Elisa, Blotting, Immunohistochemistry and Immunocytochemistry


    Some Variables to be Considered in Developing Enzyme Immunoassays


    Alternative Techniques in Immunoblotting

    Immunohistochemistry and Immunocytochemistry

    Light Microscopy Immunostaining

    Immuno-Electron Microscopy


    10. Complement



    General Properties of Complement

    Heat Instability

    Metal Ion Requirement

    Stability on Storage

    Concentration Dependence

    Proteins of the Complement System

    Activation of the Classical Pathway

    Activation of the Alternative Pathway

    The Cleavage of C5 and Assembly of the Terminal Components

    Control of the Complement System

    The Fragmentation of C3

    Complement Receptors

    Detection and Estimation of Complement Components

    Hemolytic Assays

    Table of Lysis to Sites Values

    Antigenic Assays for Complement Components

    Functional Assays and Enzyme Assays

    Detection of Activation Products

    Biological Assays

    Complement Measurement in Disease

    Genetics of the Complement System


    11. Safety

    Chemical Safety

    Risk and Safety Classification Systems

    Radiochemical Safety


    Conversion Between Units

    Protection from Radiochemical Hazards

    Microbiological Safety

    Viral Safety


    General Requirements




Product details

  • No. of pages: 256
  • Language: English
  • Published: March 7, 1994
  • Imprint: Academic Press
  • eBook ISBN: 9780080984438

About the Series Editors

B. Hames

Affiliations and Expertise

Consultant Microbiologist, Freeman Hospital, Newcastle upon Tyne, UK

D. Rickwood

Affiliations and Expertise

University of Essex, Wivenhoe Park, Colchester

About the Series Volume Editors

M. Kerr

Affiliations and Expertise

University of Dundee, Ninewells Hospital and Medical School, U.K.

Robin Thorpe

Affiliations and Expertise

National Institute for Biological Standards and Control, South Mimms, Potters Bar, U.K.

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