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Influence of cell fixatives and incubation schedule on PAP labeling of fibronectin in cultivated fibroblasts

CG Castagnino and MI Berria

A simple combined method for differential PAP labeling of fibronectin (FN) in mouse embryo fibroblast cultures was developed. Methanol-5% glacial acetic acid in dry ice-fixed cell monolayers showed mainly intracellular FN staining. Fixation with neutral paraformaldehyde before labeling, developed membrane- and extracellular matrix- associated FN. A combination of both procedures, which required incubation with primary antibody, fixation with paraformaldehyde followed by chilled acid methanol, and re-incubation with primary antibody, yielded sharp intracellular and extracellular FN labeling. The outlined methods can be readily employed in association with other staining techniques.

Volume 34, Issue 10, pp. 1331-1333, 10/01/1986
Copyright © 1986 by The Histochemical Society


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