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Detection of autoantigens by immunoblotting using a peroxidase-anti-peroxidase complex.
- Source :
-
Journal of immunological methods [J Immunol Methods] 1985 Jun 25; Vol. 80 (2), pp. 221-5. - 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 polyacrylamide 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.
Details
- Language :
- English
- ISSN :
- 0022-1759
- Volume :
- 80
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 3891864
- Full Text :
- https://doi.org/10.1016/0022-1759(85)90023-7