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Enzymatic amplification of electroimmunoprecipitates in agarose gels with horseradish peroxidase‐labeled protein A
Author(s) -
Taketa Kazuhisa
Publication year - 1983
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150040514
Subject(s) - horseradish peroxidase , agarose , conjugate , peroxidase , chemistry , chromatography , stain , microbiology and biotechnology , biochemistry , enzyme , antibody , staining , biology , mathematical analysis , mathematics , genetics , immunology
Electroimmunoprecipitates in agarose gels were amplified with protein A‐horseradish peroxidase (HRP) conjugate. The method is simple and detected quantitatively as low as 0.5 ng of human α‐fetoprotein (AFP) in electroimmunodiffusion. The alternative methods employing the second antibody, labeled initially or afterwards with HRP, failed to stain AFP immunoprecipitates. Direct application of anti‐AFP immunoglobulins‐HRP in electroimmunodiffusion produced less contrasted colored precipitates. Immunoprecipitates of other proteins were similarly visualized by the protein A‐HRP method, avoiding the use of specifically labeled antibody.