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Quantitative immunohistochemistry in the rat facial nucleus with [125I]- iodinated secondary antibodies and in situ autoradiography: non-linear binding characteristics of primary monoclonal and polyclonal antibodies

G Raivich, J Gehrmann, MB Graeber and GW Kreutzberg

Department of Neuromorphology, Max-Planck-Institute for Psychiatry, Martinsried, Germany.

Indirect immunohistochemistry is an important routine method in histology and histopathology. Here we have investigated the quantitative aspects of antibody binding to tissue sections, using a range of monoclonal and polyclonal antibodies and the regenerating rat facial nucleus as the experimental model. The in situ binding of primary antibodies was quantified using appropriate [125I]-iodinated secondary antibodies and quantitative autoradiography. The majority of primary antibodies revealed an apparently bell-shaped curve of in situ antibody binding, with the binding increasing up to a specific antibody concentration and then decreasing; similar data were also obtained with enzymatic immunohistochemistry. There was also a close correlation between the quantitative changes in antibody binding during the time course of facial nerve regeneration and those observed with enzyme histochemistry.

Volume 41, Issue 4, pp. 579-592, 04/01/1993
Copyright © 1993 by The Histochemical Society


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