Immunochemistry LabFax
- 1st Edition - March 7, 1994
- Imprint: Academic Press
- Editors: M. A. Kerr, Ali D. Hames, Robin Thorpe, D. Rickwood
- Language: English
- Paperback ISBN:9 7 8 - 0 - 1 2 - 4 1 2 1 3 7 - 9
- 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… Read more
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Request a sales quoteLABFAX 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
- Edition: 1
- Published: March 7, 1994
- No. of pages (eBook): 256
- Imprint: Academic Press
- Language: English
- Paperback ISBN: 9780124121379
- eBook ISBN: 9780080984438
MK
M. A. Kerr
Affiliations and expertise
University of Dundee, Ninewells Hospital and Medical School, U.K.AH
Ali D. Hames
Affiliations and expertise
Consultant Microbiologist, Freeman Hospital, Newcastle upon Tyne, UKRT
Robin Thorpe
Affiliations and expertise
National Institute for Biological Standards and Control, South Mimms, Potters Bar, U.K.DR
D. Rickwood
Affiliations and expertise
University of Essex, Wivenhoe Park, ColchesterRead Immunochemistry LabFax on ScienceDirect