Back to Search
Start Over
Detection of autoantigens by immunoblotting using a peroxidase-anti-peroxidase complex
- Source :
- Journal of immunological methods. 80(2)
- Publication Year :
- 1985
-
Abstract
- An immunoblotting procedure using a peroxidase-anti-peroxidase (PAP) complex was developed for the detection of autoantigens in crude mixtures by human autoimmune sera. Thymus proteins were transferred to a nitrocellulose sheet after electrophoresis in polyacrrylamide gels and probed with a 1:100 dilution of serum. The location and extent of immunoglobulin G (IgG) binding was determined by sequential reaction with: rabbit anti-human IgG, goat anti-rabbit IgG and rabbit peroxidase-anti-peroxidase complex. The peroxidase was allowed to react with chloronaphthol and low levels of autoantigen/autoantibody complex were detectable with virtual absence of background colour. The inclusion of human IgG and its pepsin-generated fragment provided a means of controlling and calibrating the blotting procedure.
- Subjects :
- Paper
Peroxidase antiperoxidase
Immunology
Autoantigens
Immunoglobulin G
snRNP Core Proteins
Immunoenzyme Techniques
chemistry.chemical_compound
Immunology and Allergy
Animals
Humans
Antigens
biology
Autoantibody
Collodion
Infant
Sodium Dodecyl Sulfate
Molecular biology
Blot
Molecular Weight
Electrophoresis
chemistry
Biochemistry
Ribonucleoproteins
biology.protein
Electrophoresis, Polyacrylamide Gel
Rabbits
Nitrocellulose
Peroxidase
Subjects
Details
- ISSN :
- 00221759
- Volume :
- 80
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Journal of immunological methods
- Accession number :
- edsair.doi.dedup.....f3c60014a65be46d58d9176e21060a36