The Quantitation of Nonspecific Staining As a Guide for Improvement of Fluorescent Antibody Conjugates

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Abstract
Summary: A technique for the precise measurement of nonspecific staining produced by conjugates labeled with fluorescein isothiocyanate has been described. Since it has given comparable results with two cell lines and in preliminary studies with a third, it is felt that data obtained by this technique can be extrapolated to a prediction of the behavior of more complex tissues when they are exposed to labeled globulins. Quantitative data have been obtained which directly relate NSS to F/P ratios at all levels and which relate specific staining to F/P ratios within certain limits.