Peroxidase anti-peroxidase method
The peroxidase-anti-peroxidase method , often abbreviated as the PAP method, is a form of immune labeling and thus serves to detect individual molecules using antibodies .
principle
The PAP method was introduced in the 1980s because it showed a lower limit of detection and less background staining than its predecessors. It was invented in 1979 by L. Sternberger. The method uses signal amplification during immunostaining by means of a sandwich of the antigen to be detected , the binding antibodies, the enzyme horseradish peroxidase (HRP, from horseradish peroxidase ) and an anti-HRP antibody. The PAP complex contains antiserum of a species and is added with the enzyme to form a complex. This complex forms a stoichiometric ratio of two IgG molecules to three enzyme molecules. It then binds to a secondary antibody, which acts as a bridge between the primary antibody and the PAP complex. This structure has the advantage that the complex remains soluble while the enzymatic activity is not influenced by the binding of the immunoglobulins .
Individual evidence
- ↑ Sternberger, L. (1979) Immunocytochemistry , 2nd ed. Wiley, New York
- ↑ Bratt Hauer, GL: The peroxidase antiperoxidase (PAP) method , in: immunocytochemical Methods and Protocols , Volume 34 of Methods in Molecular Bilogy . LC Javois, editor, 1995, Humana Press: Totowa, NJ. ISBN 978-0-89603-285-9 . Pp. 165-166.