Manual of Biological Markers of Disease

Manual of Biological Markers of Disease

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Section A: Methods of Autoantibody Detection
This Section provides well structured protocols for autoantibody detection. Step-by-step procedures are accompanied by explanatory notes and comments, clear diagrams, line illustrations and excellent photo illustrations. Extensive literature references lead the way to further background information. The methods presented were validated by more than 40 leading laboratories active in sera analysis, which indicates that these methods have been found to be practically useful and lead to consistent inter-laboratory results: consensus in autoantibody detection.
Section B: Autoantigens
This Section contains the first compilation of full, detailed information on autoantigens related to important autoimmune diseases. The chapters are all structured according to an easy-reference fixed template structure: specific detection methods, cellular localization, biochemical characteristics, function, cDNA and derived amino acid sequence, gene structure, B- and T-cell epitopes and lists of published monoclonal antibodies. The text is greatly enhanced by many beautiful schematic figures and photo illustrations. As in Section A, extensive literature references are provided. The information for this section was brought together by leading experts in their fields.
This Manual is an ideal text and reference book for bench scientists working in the field of autoimmunity, but also for rheumatologists, general (internal medicine) physicians or clinical immunologists caring for patients with autoimmune disorders.
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Product details

  • Paperback | 559 pages
  • 154.94 x 234.95 x 33.02mm | 889.04g
  • Dordrecht, Netherlands
  • English
  • Softcover reprint of the original 1st ed. 1996
  • XIV, 559 p.
  • 0792342437
  • 9780792342434

Table of contents

Section A: Methods of Autoantibody Detection.- The Editors.- 1. International cooperative activities in standardization of antinuclear antibodies.- 2. Detection of antinuclear antibodies by immunofluorescence.- 1. Introduction.- 2. The immunofluorescence assay.- 2.1 The commercial substrate.- 2.2 The culturing of HEp-2 cells.- 2.3 Fixation of the cells.- 2.4 Preparation of the serum sample.- 2.5 The conjugate antibodies.- 2.6 The incubation procedure.- 3. Interpretation of the results.- 3.1 Nuclear fluorescence patterns.- 3.2 Nucleolar fluorescence patterns.- 3.3 Spindle apparatus fluorescence patterns.- 3.4 Cytoplasmic fluorescence patterns.- 4. Solutions and suppliers.- 4.1 Solutions for culturing the cells.- 3. Counterimmunoelectrophoresis and immunodiffusion for the detection of antibodies to soluble cellular antigens.- 1. Introduction.- 2. Antigen extract preparations.- 2.1 Extraction from acetone powders.- 2.2 Whole cell extract.- 2.3 Spleen extract for Ro.- 2.4 Liver extract for aminoacyl-tRNA synthetases.- 2.5 Nuclear extract.- 2.6 Ion-exchange chromatography.- 3. Procedures.- 3.1 Immunodiffusion.- 3.2 Counterimmunoelectrophoresis (CIE).- 3.3 Staining with coomassie blue.- 3.4 Test planning and interpretation.- 4. Protein Blotting.- 1. Introduction.- 2. The antigens.- 2.1 Preparation of nuclear extracts.- 2.2 Preparation of cytoplasmic extracts.- 3. SDS polyacrylamide gel electrophoresis.- 3.1 Buffer selection.- 3.2 Power conditions.- 4. Protein blotting.- 4.1 Membrane selection.- 4.2 Blotting filter paper.- 4.3 Buffer selection.- 4.4 Semi-dry blotting.- 4.5 Tank blotting.- 4.6 Total protein staining.- 4.7 Blocking reagents.- 4.8 Serum samples.- 4.9 Immunoblot assay.- 5. Antigen appearance on the immunoblot.- 5.1 Nuclear antigens.- 5.2 Nucleolar antigens.- 5.3 Cytoplasmic antigens.- 5. Enzyme-linked immunosorbant assay in the rheumatological laboratory.- 1. Introduction.- 2. General notes.- 2.1 Solid phase.- 2.2 Conjugates.- 2.3 Substrates.- 2.4 Blocking agents.- 2.5 Antigens and antigen presentation.- 3. Assay methods.- 3.1 Solid phase ELISA.- 3.2 The chequerboard titration.- 3.3 Antigen capture (sandwich) ELISA.- 3.4 Dot immunobinding (dot blot) assay.- 4. Evaluation and improvement of the assay system.- 4.1 Evaluation.- 4.2 Improvement of the assay system.- 5. Validation of the assay system.- 6. Validation of a commercial assay system.- 7. Summary.- 8. Solutions and materials.- 8.1 Materials.- 8.2 Coating buffers.- 8.3 Dilution/washing buffer.- 8.4 Chromogenic substrate solution.- 8.5 Stop solutions.- 9. Laboratory notes on ELISA systems for the detection of antinuclear, anti-cytoplasmic and anti-phospholipid antibodies.- 9.1 Anti-nuclear antibodies.- 9.2 Anti-dsDNA.- 9.3 Anti-histone.- 9.4 Anti-Ro/SS-A.- 9.5 Anti-La.- 9.6 Anti-nRNP.- 9.7 Anti-Sm.- 9.8 Anti-ribosomal RNP (P protein).- 9.9 Anti-Sc170 (Topoisomerase 1).- 9.10 Anti-Ki.- 9.11 Anti-Jo-1.- 9.12 Anti-Cardiolipin.- 9.13 Anti-Centromere.- 6. Immunoprecipitation of labelled proteins.- 1. Introduction.- 2. Protein immunoprecipitation procedures.- 2.1 Preparation of 35S-methionine radiolabeled extract.- 2.2 Preparation of the gels.- 3. Interpretation.- 3.1 Sm and U1RNP.- 3.2 SS-B/La and SS-A/Ro.- 3.3 PCNA, Jo-1, rRNP and Ku.- 4. Materials.- 4.1 Reagents and equipment.- 5. Solutions.- 5.1 Tissue culture media.- 5.2 Acrylamide solutions and electrophoresis buffers.- 5.3 Other buffers and solutions.- 7. Analysis of autoimmune sera by immunoprecipitation of cellular RNPs.- 1. Introduction.- 2. Preparation of cellular extracts.- 2.1 Radioactive labeling of RNAs.- 2.2 Cell fractionation.- 3. Immunoprecipitation.- 3.1 Binding of antibodies to agarose beads and immunoprecipitation of RNP complexes.- 4. RNA gel electrophoresis.- 4.1 10% acrylamide/8.3 M urea gel.- 4.2 6-15% gradient acrylamide gels.- 4.3 Silver staining procedure.- 5. Reagents and suppliers.- 8. Measurement of antibodies to DNA.- 1. Introduction.- 2. Procedures.- 2.1 ELISA.- 2.2 IFT on Crithidia luciliae.- 2.3 PEG assay.- 2.4 Farr assay.- 3. Reagents and solutions.- 3.1 ELISA.- 3.2 IFT on Crithidia luciliae.- 3.3 PEG assay.- 3.4 Farr assay.- 9. Methods to detect autoantibodies to neutrophilic granulocytes.- 1. Introduction.- 2. Demonstration of neutrophil-reactive autoantibodies by IIF.- 2.1 Steps in the procedure.- 2.2 Solutions.- 2.3 Interpretation, artifacts, control cells.- 3. EIA procedures.- 3.1 Preparation of ?-granules as described by Borregaard et al..- 3.2 EIA for quantification of ANCA using ?-granules.- 3.3 EIA for quantification of Proteinase-3 (PR-3) ANCA.- 3.4 EIA for quantification of myeloperoxidase (MPO) ANCA.- 10. Detection of antiperinuclear factor and antikeratin antibodies.- 1. Introduction.- 2. Detection of the APF.- 2.1 Antigen substrate and their donors.- 2.2 Preparation of buccal mucosa cells for immunofluorescence.- 2.3 Serum dilution and immunofluorescence procedure.- 2.4 Criteria for positivity and reproducibility of the test.- 2.5 Reagents and solutions.- 3. Detection of antikeratin antibodies (AKA).- 3.1 Antigen substrate.- 3.2 Serum dilution and immunofluorescence procedure.- 3.3 Criteria for positivity of the AKA test.- 3.4 Reagents and solutions.- 11. Standards and reference preparations.- 1. Introduction.- 2. Standards.- 3. Reference preparations.- 4. How to use a standard.- 5. Why are standards often not used?.- 6. Examples of the use of standards.- 7. Characteristics of standards and reference preparations.- 8. How to obtain the standards?.- Section B: Autoantigens.- The Editors.- 1. Autoantigens in Rheumatoid Arthritis (RA).- 1. RF (rheumatoid factor).- 2. APF (antiperinuclear factor).- 3. RA-33 (hnRNP-A2).- 4. Collagen II.- 2. Autoantigens in Systemic Lupus Erythematosus (SLE).- 1. DNA.- 2. Histones.- 3. Ubiquitin.- 4. Sm.- 5. Ribosomal RNP.- 6. Ku.- 7. PCNA.- 8. Phospholipids.- 3. Autoantigens in SLE-overlap syndroms.- 1. The U1 snRNP complex.- 4. Autoantigens in Sjoegren's syndrome.- 1. Ro/SS-A.- 2. La/SS-B.- 5. Autoantigens in Scleroderma.- 1. DNA topoisomerase 1.- 2. Centromere Proteins.- 3. Fibrillarin.- 4. PM/Scl.- 6. Autoantigens in Myositis.- 1. Transfer RNA synthetases.- 7. Autoantigens in Vasculitis.- 1. Proteinase 3 (c-ANCA).- 2. Myeloperoxidase (MPO).- 8. Autoantigens in other diseases.- 1. Mitochondrial autoantigens.- 2. P80 coilin.
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