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  • Produktbild: Methods of Protein and Nucleic Acid Research
  • Produktbild: Methods of Protein and Nucleic Acid Research

Methods of Protein and Nucleic Acid Research 2 Immunoelectrophoresis Application of Radioisotopes

Fr. 72.90

inkl. gesetzl. MwSt., Versandkostenfrei

Beschreibung

Produktdetails

Einband

Taschenbuch

Erscheinungsdatum

29.04.2012

Verlag

Springer Berlin

Seitenzahl

206

Maße (L/B/H)

24.4/17/1.3 cm

Gewicht

389 g

Auflage

Softcover reprint of the original 1st edition 1984

Sprache

Englisch

ISBN

978-3-642-87490-1

Beschreibung

Produktdetails

Einband

Taschenbuch

Erscheinungsdatum

29.04.2012

Verlag

Springer Berlin

Seitenzahl

206

Maße (L/B/H)

24.4/17/1.3 cm

Gewicht

389 g

Auflage

Softcover reprint of the original 1st edition 1984

Sprache

Englisch

ISBN

978-3-642-87490-1

Herstelleradresse

Springer-Verlag GmbH
Tiergartenstr. 17
69121 Heidelberg
DE

Email: ProductSafety@springernature.com

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  • Produktbild: Methods of Protein and Nucleic Acid Research
  • Produktbild: Methods of Protein and Nucleic Acid Research
  • 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 Counting of the Eluates.- 2.12 Autoradiography.- 2.12.1 Drying of PAAG Slabs.- 2.13 Indirect Autoradiography.- 2.14 Fluorography.- 2.14.1 Fluorography of Plates After TLC.- 2.14.2 Fluorography of PAAG Slabs.- 2.15 Increase in X-ray Film Sensitivity by Prior Exposure to Light.- 2.16 Quantitative Counting.- 2.17 Counting of Dual Label in PAAG.- 2.18 Scanning Plates with Endwindow Counters (Berthold Company).- 2.18.1 Scanning Instrument.- 2.18.2 Berthold LB 2832 Linear Analyzer.- 2.18.3 Photography of Radioactive Spots on Plates Using a Spark Chamber (Berthold LB 292).- 2.19 Detection of Radioactivity in a Flowing Liquid.- 3 Labelling of the Proteins.- 3.1 Iodination with Chloramine T.- 3.2 Iodination in the Presence of H2O2 and Lactoperoxidase.- 3.3 Addition of Special Iodinated Reagents.- 3.4 Iodination with Na125I in the Presence of ICl.- 3.5 Labelling with 14C and 3H Using Formaldehyde and Borohydride.- 3.6 Enzymatic Methods.- 4 Labelling of Nucleic Acids.- 4.1 Chemical Reactions.- 4.1.1 Methylation with Labelled Dimethylsulfate (DMS).- 4.1.2 Incorporation of 3H from Tritiated Water in the Presence of Sodium Metabisulfite (Na2S2O5).- 4.1.3 Slow Isotope Exchange of Hydrogen.- 4.1.4 Labelling the 3?-Terminal Ribonucleotide: The Method of Randerath.- 4.1.5 Complexing Denatured DNA with Labelled N-hydroxymethylene Aminoacid.- 4.1.6 Iodination of Nucleic Acids.- 4.2 Enzymatic Reactions.- 4.2.1 T4-Polynucleotide Kinase.- 4.2.2 Terminal Deoxynucleotide Transferase.- 4.2.3 T4 RNA-Ligase.- 4.2.4 DNA-Polymerase I from E. coli (Kornberg Enzyme).- 5 Labelling of Proteins and Nucleic Acids in Vivo.- 5.1 Higher Animals.- 5.2 Cell Cultures from Higher Organisms.- 5.3 Microorganisms.- 6 Determination of Concentrations by Isotope Dilution.- 7 Radioimmune Methods.- 7.1 Immunoprecipitation and Adsorption from Solution.- 7.1.1 The System of Two Antibodies.- 7.1.2 The Use of Staphylococcus aureus and of Sepharose-linked Protein A.- 7.1.3 Radioactively Labelled Antibodies.- 7.1.4 Radioactive Protein A.- 7.1.5 Immunoprecipitation System with Avidin-Biotin Complex.- 7.2 Competitive Radioimmunoassay (RIA).- 7.2.1 RIA with Precipitation of the Immune Complexes.- 7.2.2 RIA with Filtration.- 7.2.3 Solid-phase RIA.- 7.3 Enzymic Immune Methods.- Some Additional Remarks on the Use of Radioactive Isotopes.- References to Part I.- References to Part II.