Holiday book sale: Save up to 30% on print and eBooks. No promo code needed.
Save up to 30% on print and eBooks.
1st Edition - March 7, 1994
Editors: Ali D. Hames, M. A. Kerr, Robin Thorpe, D. Rickwood eBook ISBN:
9 7 8 - 0 - 0 8 - 0 9 8 4 4 3 - 8
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…
Save 50% on book bundles
Immediately download your ebook while waiting for your print delivery. No promo code is needed.
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. Contributors Abbreviations 1. Plasma Proteins Plasma and Serum Anticoagulants Properties of Commonly Used Anticoagulants (Table 1) Serum 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 References 2. Properties of Immunoglobulins Introduction 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) References 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 References 4. Producing Antibodies Polyclonal Antisera Preparation of Immunogen Haptenization of Peptides Adjuvants 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 Cloning Production of Monoclonal Antibodies Production of Human Monoclonal Antibodies by Lymphocyte Transformation rDNA-Produced Antibodies Spliced Immunoglobulin Molecules Recombinant DNA-Derived Immunoglobulin Fragments References 5. Purification and Fragmentation of Immunoglobulins General Properties of Antibodies Stability 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 References 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) Radioimmunoassay Buffer Assay Components Data Analysis Further Optimization Immunoradiometric Assay Assay Components Data Analysis Assay Optimization References 7. Enzyme-Conjugated Antibodies: Preparation and Use Introduction Enhanced Enzyme Immunodetection Systems Enzymes Commonly Used in Immunoconjugates Horseradish Peroxidase Alkaline Phosphatase ß-Galactosidase Glucose Oxidase Urease 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 References 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 General Properties of Biotin Properties of Avidin Properties of Streptavidin Colloidal Gold Derivatives Properties of Colloidal Gold Conjugates References 9. Common Immunological Techniques: Elisa, Blotting, Immunohistochemistry and Immunocytochemistry Elisa Some Variables to be Considered in Developing Enzyme Immunoassays Immunoblotting Alternative Techniques in Immunoblotting Immunohistochemistry and Immunocytochemistry Light Microscopy Immunostaining Immuno-Electron Microscopy References 10. Complement Introduction Nomenclature 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 References 11. Safety Chemical Safety Risk and Safety Classification Systems Radiochemical Safety Definitions Conversion Between Units Protection from Radiochemical Hazards Microbiological Safety Viral Safety Introduction General Requirements Centrifugation References Index
eBook ISBN: 9780080984438
Ali D. Hames
Affiliations and expertise
Consultant Microbiologist, Freeman Hospital, Newcastle upon Tyne, UK M. A. 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. D. Rickwood
Affiliations and expertise
University of Essex, Wivenhoe Park, Colchester