Methods of Protein and Nucleic Acid Research: 2 Immunoelectrophoresis Application of Radioisotopes (Paperback, Softcover Repri)

I Immunoelectrophoresis.- 1 Fundamentals of Immunochemistry.- 1.1 The Immune System of Higher Organisms.- 1.1.1 Blood Components.- 1.1.2 Phagocytosis, Complement, Cytointoxication.- 1.1.3 Leucocytes.- 1.1.4 Antibodies.- 1.1.5 The Lymphatic System and the Spleen.- 1.1.6 Decay of the Immune Response.- 1.1.7 Kinetics of Immunization.- 1.1.8 Generation of the Immunological Arsenal.- 1.2 Structure of the Immunoglobulins.- 1.2.1 General Description of Structure. Classes of Immunoglobulins.- 1.2.2 Antigen-binding Sites.- 1.2.3 Ambiguity of the Immune Response.- 1.2.4 Synthesis and Assembly of Immunoglobulin Molecules.- 1.3 Myeloma and Hybridoma Cells.- 1.4 Preparation of Immune Serum and Purification of Antibodies.- 1.4.1 Immunization of the Rabbit.- 1.4.2 Preparation of Antiserum.- 1.4.3 Purification of Immunoglobulin G (IgG).- 1.4.4 The Use of Immunosorbents for Purification of Antibodies.- 1.5 Production of Monoclonal Antibodies Using Hybridoma.- 1.6 Commercially Available Antibodies.- 1.7 Detection and Estimation of Antibody Concentration.- 1.7.1 Ring Test.- 1.7.2 Determination of the Ratio of Equivalence.- 1.7.3 Hemagglutination and Hemolysis.- 1.7.4 Complement Binding Reaction.- 1.7.5 Ouchterlony's Method.- 2 The Use of Immune Methods to Detect Protein Zones Following Conventional Electrophoresis and Electrofocusing.- 2.1 Immunochemical Detection of Antigens in Gels.- 2.2 Preparation of Immune Replicas.- 2.2.1 Agarose Replica.- 2.2.2 Diazopaper Replica.- 2.2.3 Nitrocellulose Replica.- 3 Immunoelectrophoresis According to Grabar and Williams.- 3.1 Running the Experiment.- 4 Laurell Rocket Immunoelectrophoresis.- 4.1 Principle of the Method.- 4.2 Running the Experiment. Description of the Underlying Processes.- 5 Crossed Immunoelectrophoresis.- 5.1 Method of Clarke and Freeman.- 5.2 Tandem Crossed Immunoelectrophoresis.- 5.3 Methods of Fractionation in the First Dimension.- 5.4 Technique of Overlaying Agarose Gel with PAAG.- 5.5 The Use of an Intermediate Gel.- 5.6 Crossed Immunoaffinity Electrophoresis.- 5.7 Fused Rocket Immunoelectrophoresis.- II The Use of Radioisotopes.- 1 Isotopes, Scintillators, and Scintillation Counters.- 1.1 Radioactive Isotopes.- 1.1.1 Excess of Neutrons.- 1.1.2 Shortage of Neutrons.- 1.1.3 Rate of Radioactive Decay.- 1.1.4 Total Radioactivity of the Sample.- 1.1.5 Specific Activity (SA).- 1.1.6 Carrier-free Radioisotopes.- 1.2 Scintillators.- 1.2.1 Liquid Scintillators.- 1.2.2 The Spectrum of Light Pulse Intensities.- 1.2.3 Counting Efficiency.- 1.2.4 Quenching.- 1.2.5 ?-Radiation Detection. Solid Scintillators.- 1.3 Liquid Scintillation Counters. Principle of Operation.- 1.3.1 Photomultiplier.- 1.3.2 Amplifiers (Linear and Logarithmic).- 1.3.3 Logarithmic Voltage Amplifier.- 1.3.4 Pulse-Height Discriminators. "Thresholds".- 1.3.5 Background Noise.- 1.3.6 Coincidence Circuit.- 1.3.7 Scalers.- 1.3.8 Accuracy of Counting.- 1.3.9 Chemiluminescence.- 1.3.10 Photoluminescence.- 1.3.11 Counting Vials.- 1.3.12 Cerenkov Counting.- 1.4 Adjustment (Tuning) of Liquid Scintillation Counters.- 1.4.1 Adjustment of One Channel for Single Isotope Counting.- 1.4.2 Adjustment of Two Channels for Dual Isotope Counting.- 1.4.3 Quench Correction for Single Isotope Counting.- 1.4.4 Quench Correction for Dual Isotope Counting.- 1.4.5 Adjustment of Two Channels for Dual Isotope Counting (3H and 14C) in a Counter with a Linear Amplifier.- 1.4.6 Dual Label Counting of 3H and 125I in a Liquid Scintillation Counter.- 2 Preparation and Counting of Labelled Samples.- 2.1 Samples as Aqueous Solutions.- 2.2 Dioxane-based Scintillators.- 2.3 Toluene- and Xylene-based Scintillators.- 2.4 Solubilization of Biological Specimens.- 2.5 Counting on Filters.- 2.6 Counting Efficiency. The Recovery of Radioactivity.- 2.7 Counting of Powders in Suspension.- 2.8 Counting After TLC and Gel Electrophoresis.- 2.9 Digestion of PAAG with 30% H2O2.- 2.10 Solubilization of PAAG without H2O2.- 2.11 Elution of Proteins and Nucleic Acids from Gels and