Specificity controls for immunocytochemical methods

  • Burry R
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SUMMARY Immunocytochemistry is used for antibody localization of proteins in cells and tissues. The specificity of the results depends on two independent criteria: the specificity of the antibody and of the method used. The antibody specificity is best determined by immu-noblot and or immunoprecipitation. Absorption of the antibody with a protein does not determine that the antibody would have bound to the same protein in the tissue, and therefore is not a good control for antibody specificity. The specificity of the method is best determined by both a negative control, replacing the primary antibody with serum, and a positive control, using the antibody with cells known to contain the protein. With the in-creasing use of immunocytochemistry, it is important to be aware of the appropriate con-trols needed to show specificity of the labeling. (J Histochem Cytochem 48:163–165, 2000)

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  • Richard W. Burry

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