Summary
A modified method is described for the rapid production of peroxidase-antiperoxidase complexes to be used in immunocytochemistry. In this method anti-peroxidase antibodies are precipitated from crude serum with peroxidase at equivalence and subsequently resolubilized at low pH with excess peroxidase. The complexes are isolated from unbound immunoglobulin and peroxidase by gelfiltration. The method combines the advantages of both previously described preparation procedures. The resulting PAP-complex, when tested in indirect immunocytochemistry, is comparable to that obtained in established preparation procedures.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Avrameas S (1969) Coupling of enzymes to proteins with glutaraldehyde. Use of conjugates for the detection of antigens and antibodies. Immunochemistry 6:43
Bosman FT, Nieuwenhuijzen Kruseman AC (1979) Clinical application of the enzyme labeled antibody method. Immunoperoxidase methods in diagnostic histopathology. J. Histochem Cytochem 27:1140–1147
Mason TE, Phifer RF, Spicer SS, Swallow RA, Dreskin RB (1969) An immunoglobulin-enzymebridge method for localizing tissue antigens. J Histochem Cytochem 17:563–569
Mason DY, Sammons R (1978) Rapid preparation of peroxidase — antiperoxidase complexes for immunocytochemical use. J Immunol Methods 20:317–324
Mason DY, Sammons R (1979) The labeled antigen method of immunoenzymatic staining. J Histochem Cytochem 27:832–840
Nakane PK, Kawaoi A (1974) Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem 22:1084–1091
Nieuwenhuijzen Kruseman AC, Knijnenburg G, Brutel de la Riviere G, Bosman F (1978) Morphology and immunohistochemically-defined endocrine function of pancreatic islet cell tumours. Histopathology 2:389–399
Sternberger LA (1978) Immunocytochemistry. John Wiley & Sons, New York
Sternberger LA, Cuculis JJ (1969) Method of enzymatic intensification of the immunocytochemical reaction without use of labeled antibodies. J Histochem Cytochem 17:190
Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method for immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-anti-horseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333
Taylor CR (1978) Immunoperoxidase techniques. Practical and theoretical aspects. Arch Pathol Lab Med 102:113–121
Wilson MB, Nakane PK (1978) Recent developments in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies. In: Knapp W, Holubar K, Wick G (eds) Immunofluorescence and related staining techniques. Elsevier/North Holland, Amsterdam New York pp 215–221
Author information
Authors and Affiliations
Additional information
Supported by a grant from the Queen Wilhelmina Cancer Foundation (grant no. PA 77/52)
Rights and permissions
About this article
Cite this article
Bosman, F.T., Cramer-Knijnenburg, G. & van Bergen Henegouw, J. A simplified method for the rapid preparation of peroxidase-anti-peroxidase (PAP) complexes. Histochemistry 67, 243–248 (1980). https://doi.org/10.1007/BF00692758
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00692758