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Wiley, Diagnostic Cytopathology, 1(10), p. 82-85, 1994

DOI: 10.1002/dc.2840100120

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Methodological aspects of the immunostaining of proliferating cell nuclear antigen (PCNA) in cytospin preparations of MCF-7 cell line

This paper was not found in any repository, but could be made available legally by the author.
This paper was not found in any repository, but could be made available legally by the author.

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Abstract

Cytospins of a human breast cancer cell line (MCF‐7) were studied for the expression of PCNA, a cell cycle‐related protein, using a variety of fixation and immunostaining procedures. The best fixative for PCNA was found to be buffered formaldehyde solution at 4°C followed by methanol at 20°C, whereas alcoholic fixatives decreased greatly the PCNA immunoreactivity. Air‐drying procedures of cytospins prior to and after fixation greatly undermined the PCNA immunostaining. A modified immunoperoxidase method provided a stronger staining of the PCNA‐reactive cells than the alkaline phosphatase anti‐alkaline phosphatase (APAAP) technique. PCNA immunoreactivity could be maintained up to 2 mo, putting slides in methanol at − 2°C. In conclusion, our report indicates that PCNA is a labile antigen, which may critically be affected by temperature and air‐drying procedures