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Manual of biological markers of disease

Author: W J van Venrooij; R N Maini
Publisher: Dordrecht ; Boston : Kluwer Academic Publishers, ©1996.
Edition/Format:   Print book : EnglishView all editions and formats
Summary:

Section A: Methods of Autoantibody Detection This Section provides well structured protocols for autoantibody detection.

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Document Type: Book
All Authors / Contributors: W J van Venrooij; R N Maini
ISBN: 0792342429 9780792342427 0792342437 9780792342434
OCLC Number: 35236040
Description: 1 volume (various pagings) : illustrations ; 24 cm
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.
Responsibility: edited by W.J. van Venrooij and R.N. Maini.

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