PMID: 7542012Mar 1, 1995Paper

The identification of proliferating cell nuclear antigen (PCNA) on rat tissue cryosections and its application to double immunostaining with other markers

Archives of Histology and Cytology
T EzakiK Matsuno

Abstract

Proliferating cell nuclear antigen (PCNA) in a useful marker for monitoring cell proliferation in most species. The immunostaining of PCNA on tissue cryosections, however, has been hampered by a loss of PCNA immunoreactivity during the staining process. The need for both identifying the actual phenotypes of proliferating cells and differentiating them from other tissue components prompted us to establish reliable techniques for PCNA immunostaining on cryosections and to apply these to double immunostainings with other markers. We tested various fixing conditions for rat tissue cryosections and the effect derived from unmasking with pepsin digestion to restore PCNA immunoreactivity after fixation. For single immunostainings, the unmasking was effective in most fixing conditions tested. Particularly, 4% paraformaldehyde/0.05% glutaraldehyde fixation followed by 0.001% pepsin digestion resulted in the strongest immunoreactivity for PCNA and the best morphology, and was the first choice for double immunostainings with relatively stable second antigens. Alternatively, periodate-lysine-paraformaldehyde fixation was also applicable to other second antigens which are labile to the former treatment. These techniques can serve in the col...Continue Reading

Citations

Jun 8, 2001·The American Journal of Pathology·T EzakiD M McDonald
Jun 6, 2000·Micron : the International Research and Review Journal for Microscopy·T Ezaki
Jul 8, 1999·The American Journal of Pathology·T J MurphyD M McDonald
Jan 22, 2002·The Journal of Immunology : Official Journal of the American Association of Immunologists·Koichi SaitoMasayuki Miyasaka
Jul 9, 2003·The Journal of Immunology : Official Journal of the American Association of Immunologists·Daisuke NagakuboMasayuki Miyasaka
Aug 6, 2019·Clinical Anatomy : Official Journal of the American Association of Clinical Anatomists & the British Association of Clinical Anatomists·Soosai Manickam AmirthamElizabeth Vinod
Mar 9, 2021·Biological Trace Element Research·Ziqiang ZhangYumei Liu
Dec 10, 2008·Journal of Immunological Methods·Fabio D'AmicoFranca Stivala

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