Summary
Mouse antibodies are increasingly used as primary antibodies for immunocytochemistry as more mouse monoclonal antibodies are being produced. The localisation of these antibodies by the PAP technique requires mouse antiperoxidase antibody. A monoclonal antiperoxidase would obviate the limitations of production of a polyclonal mouse antiperoxidase. This paper describes the development of a mouse hybridoma producing such an antibody (MAP A6-2) and the use of this antibody to localise a number of mouse primary antibodies by the PAP technique for both light and electron microscopy. The antibodies localised include monoclonal antienkephalin and antityrosine hydroxylase. MAP A6-2 had a higher affinity in immuno-diffusion experiments and gives slightly better staining with an horse radish peroxidase of a different type from that used for immunisation. Staining was optimum with horse radish peroxidase type X whereas horse radish peroxidase type VI was used for immunisation. Also described is the production of a HAT sensitive variant cell line allowing the possibility of using this hybridoma as a parent cell line for the production of hybrid hybridomas secreting bi-specifie antibodies.
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Cuello AC, Milstein C, Couture R, Wright B, Priestley JV, Jarvis J (1984a) Characterisation and immunocytochemical applications of monoclonal antibodies against enkephalins. J Histochem Cytochem 32:947–957
Cuello AC, Milstein C, Wright B, Bramwell S, Priestley JV, Jarvis J (1984b) Development and application of a monoclonal rat peroxidase antiperoxidase (PAP) immunocytochemical reagent. Histochemistry 80:257–261
Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of pre-defined specificity. Nature 256:495–497
Kohler G, Milstein C (1976) Derivation of specific antibody producing tissue culture and tumour lines by cell fusion. Eur J Immol 6:511–519
Kohler G, Howe SC, Milstein C (1976) Fusion between immunoglobulin secreting and non secreting myeloma cell lines. Eur J Immunol 6:292
MacMillan FM, Sidebottom E, Cuello AC (1984b) A monoclonal antibody recognising an intracellular antigen expressed differentially in neural cell types. Trajabos del Instituto Cajal, LXXV Suppl p 89
Mason DY, Cordell JL, Abdulaziz Z, Naiem M, Bordenave G (1982) Preparation of peroxidase:antiperoxidase (PAP) complexes for immunohistological labeling of monoclonal antibodies. J Histochem Cytochem 30:114–122
Milstein C, Cuello AC (1983) Hybrid hybridomas and their use in immunohistochemistry. Nature 304:537–540
Milstein C, Cuello AC (1984) Hybrid hybridomas and the production of bi-specific monoclonal antibodies. Immunol Today 5:299–304
Priestley JV, Cuello AC (1983) Electron microscopic immunocytochemistry: Central nervous system transmitters and transmitter markers. In: Cuello AC (ed) Immunocytochemistry. John Wiley, Chichester, pp 273–322
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) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333
Vaitukaitis J, Robbins JB, Nieschlog E, Ross GT (1971) A method for producing antisera with small doses of immunogen. J Clin Endocrinol Metab 33:988
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Semenenko, F.M., Bramwell, S., Sidebottom, E. et al. Development of a mouse antiperoxidase secreting hybridoma for use in the production of a mouse PAP complex for immunocytochemistry and as a parent cell line in the development of hybrid hybridomas. Histochemistry 83, 405–408 (1985). https://doi.org/10.1007/BF00509200
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DOI: https://doi.org/10.1007/BF00509200