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Practical suggestions for successful immunoenzyme double-staining experiments

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Summary

Many methodologies exist to perform an immunoenzyme double staining. Hence, the practical problem arises as to which of these methods is optimal for one's own experimental design. A process of selection is described which is derived from our own practical experience. First, a general strategy is outlined for the handling of tissue sections to be used for multiple staining methods. Secondly, the selection of an appropriate immunoenzyme double-staining concept is made using a flow chart. Thereafter we give criteria for the definitive selection of an immunoenzyme double-staining protocol based on the characteristics of the tissue or cell type under study. Particular attention is given to the selection of appropriate detection systems, applying enzymes or gold particles, and good contrasting colour combinations. The problems of visualizing co-localization using immunoenzyme double staining are dealt with, and suggestions are made to adapt the method, if necessary, in order to optimize it.

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  1. Academic Medical Center, Cardiovascular Pathology Unit, Meibergdreef 9, NL-1105 AZ, Amsterdam, The Netherlands

    Chris M. van der Loos & Anton E. Becker

  2. Department of Pathology, University Hospital St Rafael, Kapucijnenvoer 33, B -3000, Leuven, Belgium

    Joost J. van den Oord

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  1. Chris M. van der Loos
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  2. Anton E. Becker
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This paper (in modified form) is part of the thesis of C. M. van der Loos: Free University Press 1992, Amsterdam, The Netherlands (ISBN 90-5383-081-2).

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van der Loos, C.M., Becker, A.E. & van den Oord, J.J. Practical suggestions for successful immunoenzyme double-staining experiments. Histochem J 25, 1–13 (1993). https://doi.org/10.1007/BF00161039

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