Summary
Monoclonal antibodies are being increasingly used in immunocytochemistry but their localisation by the peroxidase antiperoxidase (PAP) procedure requires the use of rat or mouse PAP. In this paper we describe the development and application of a monoclonal rat PAP. This reagent has been used successfully for immunocytochemistry at light and electron microscopy level in combination with rat monoclonal antibodies against serotonin (5-HT), substance P and somatostatin. The monoclonal rat PAP has several advantages over conventional polyclonal rat PAP and is likely to be a valuable developing reagent in immunocytochemistry using rat monoclonal antibodies.
Similar content being viewed by others
References
Consolazione A, Milstein C, Wright B, Cuello AC (1981) Immunocytochemical detection of serotonin with monoclonal antibodies. J Histochem Cytochem 29:1425–1430
Cuello AC, Galfre G, Milstein C (1979) Detection of substance P in the central nervous system by a monoclonal antibody. Proc Natl Acad Sci USA 76:3532–3536
Cuello AC, Priestley JV, Milstein C (1982) Immunocytochemistry with internally labelled monoclonal antibodies. Proc Natl Acad Sci USA 79:665–669
Galfre G, Milstein C (1981) Preparation of monoclonal antibodies, strategies and procedures. Methods Enzymol 73:3–46
Galfre G, Milstein C, Wright B (1979) Rat x rat hybrid myelomas and a monoclonal anti-Fd portion of mouse IgG. Nature 277:131–133
Howe PRC, Cuello ACC, Costa M, Furness JB (1982) Improved immunohistochemical visualisation of central serotonin nerves after loading with 5,7-dihydroxytryptamine. Neurosci Lett 29:1–6
Kilmartin JD, Wright B, Milstein C (1982) Rat monoclonal antitubulin antibody derived by using a new nonsecreting rat cell line. J Cell Biol 93:576–582
Mason DY, Cordell J, Abdulaziz Z, Naiem M, Bordenave G (1982) Preparation of peroxidase: antiperoxidase (PAP) complexes for immunohistological labeling of monoclonal antibodies. J Histochem Cytochem 30:1114–1122
Milstein C, Cuello AC (1983) Hybrid hybridomas and their use in immunohistochemistry. Nature 305:537–540
Priestley JV, Cuello AC (1983) Electron microscopic immunocytochemistry for CNS trasmitters and transmitter markers. In: Cuello AC (ed) Immunohistochemistry. John Wiley & Sons, London, pp 273–322
Priestley JV, Somogyi P, Cuello AC (1981) Neurotransmitter specific projection neurons revealed by combining PAP immunohistochemistry with retrograde transport of HRP. Brain Res 220:231–240
Priestley JV, Somogyi P, Cuello AC (1982) Immunocytochemical localisation of substance P in the spinal trigeminal nucleus of the rat: a light and electron microscopic study. J Comp Neurol 221:31–49
Sternberger LA (1979) Immunocytochemistry. 2nd edn. John Wiley, New York
Sternberger LA, Hardy PH, Cuculis JJ, Meyer HG (1970) The unlabelled antibody-enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333
Vandesande F (1983) Peroxidase-antiperoxidase techniques. In: Cuello AC (ed) Immunohistochemistry. John Wiley & Sons, London, pp 101–119
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Cuello, A.C., Milstein, C., Wright, B. et al. Development and application of a monoclonal rat peroxidase antiperoxidase (PAP) immunocytochemical reagent. Histochemistry 80, 257–261 (1984). https://doi.org/10.1007/BF00495774
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00495774