Antibody for detecting p53 protein by immunohistochemistry in normal tissues

F. Pezzella, K. Micklem, H. Turley, K. Pulford, M. Jones, S. Kocialkowski, D. Delia, A. Aiello, R. Bicknell, K. Smith, A. L. Harris, K. C. Gatter, D. Y. Mason

Research output: Contribution to journalArticle

Abstract

Aims-To establish whether PAb248 recognises human p53 as weli as murine p53 and if so, to determine its distribution in normal tissues. Methods-The ability of PAb248 to recognise human p53 was established by analysis of the human osteosarcoma derived Saos-2 celil line, which lacks the p53 gene, before and after transfection with p53 cDNA, using western blotting and immunoprecipitation. Immunostaining on normal tissues and cell lines was carried out using an immunoperoxidase technique. The two anti-p53 antibodies PAb 240 and DO-7 were used as controls. Results-The anti-p53 PAb248 monoclonal antibody stained the Saos-2 cell line after, but not before, transfection with p53 cDNA. Both western blots and immunoprecipitations performed with this antibody revealed a 53 000 molecular weight band. With immunostaining, this antibody detects p53 protein in most lymphoid and human epithelial cells in a cytoplasmic-perinuclear localisation that has not been described before. In the same tissues nuclear staining could be seen in a few scattered cells using the PAb240 antibody. The topographical distribution of wild type p53 was not related to proliferating areas but, rather, to short-lived populations of cells. Conclusions-Immunostaining of wild type p53 is demonstrable not only in its nuclear form using antibody PAb240 but also in its common cytoplasmic- perinuclear localisation in normal tissues using the PAb248 monoclonal antibody. This opens up new possibilities for its study in both physiological and pathological conditions.

Original languageEnglish
Pages (from-to)592-596
Number of pages5
JournalJournal of Clinical Pathology
Volume47
Issue number7
DOIs
Publication statusPublished - 1994

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Immunohistochemistry
Antibodies
Immunoprecipitation
Transfection
Proteins
Complementary DNA
Western Blotting
Monoclonal Antibodies
Cell Line
Normal Distribution
p53 Genes
Osteosarcoma
Immunoenzyme Techniques
Anti-Idiotypic Antibodies
Molecular Weight
Epithelial Cells
Staining and Labeling
Population

ASJC Scopus subject areas

  • Health Policy
  • Pathology and Forensic Medicine

Cite this

Pezzella, F., Micklem, K., Turley, H., Pulford, K., Jones, M., Kocialkowski, S., ... Mason, D. Y. (1994). Antibody for detecting p53 protein by immunohistochemistry in normal tissues. Journal of Clinical Pathology, 47(7), 592-596. https://doi.org/10.1136/jcp.47.7.592

Antibody for detecting p53 protein by immunohistochemistry in normal tissues. / Pezzella, F.; Micklem, K.; Turley, H.; Pulford, K.; Jones, M.; Kocialkowski, S.; Delia, D.; Aiello, A.; Bicknell, R.; Smith, K.; Harris, A. L.; Gatter, K. C.; Mason, D. Y.

In: Journal of Clinical Pathology, Vol. 47, No. 7, 1994, p. 592-596.

Research output: Contribution to journalArticle

Pezzella, F, Micklem, K, Turley, H, Pulford, K, Jones, M, Kocialkowski, S, Delia, D, Aiello, A, Bicknell, R, Smith, K, Harris, AL, Gatter, KC & Mason, DY 1994, 'Antibody for detecting p53 protein by immunohistochemistry in normal tissues', Journal of Clinical Pathology, vol. 47, no. 7, pp. 592-596. https://doi.org/10.1136/jcp.47.7.592
Pezzella F, Micklem K, Turley H, Pulford K, Jones M, Kocialkowski S et al. Antibody for detecting p53 protein by immunohistochemistry in normal tissues. Journal of Clinical Pathology. 1994;47(7):592-596. https://doi.org/10.1136/jcp.47.7.592
Pezzella, F. ; Micklem, K. ; Turley, H. ; Pulford, K. ; Jones, M. ; Kocialkowski, S. ; Delia, D. ; Aiello, A. ; Bicknell, R. ; Smith, K. ; Harris, A. L. ; Gatter, K. C. ; Mason, D. Y. / Antibody for detecting p53 protein by immunohistochemistry in normal tissues. In: Journal of Clinical Pathology. 1994 ; Vol. 47, No. 7. pp. 592-596.
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abstract = "Aims-To establish whether PAb248 recognises human p53 as weli as murine p53 and if so, to determine its distribution in normal tissues. Methods-The ability of PAb248 to recognise human p53 was established by analysis of the human osteosarcoma derived Saos-2 celil line, which lacks the p53 gene, before and after transfection with p53 cDNA, using western blotting and immunoprecipitation. Immunostaining on normal tissues and cell lines was carried out using an immunoperoxidase technique. The two anti-p53 antibodies PAb 240 and DO-7 were used as controls. Results-The anti-p53 PAb248 monoclonal antibody stained the Saos-2 cell line after, but not before, transfection with p53 cDNA. Both western blots and immunoprecipitations performed with this antibody revealed a 53 000 molecular weight band. With immunostaining, this antibody detects p53 protein in most lymphoid and human epithelial cells in a cytoplasmic-perinuclear localisation that has not been described before. In the same tissues nuclear staining could be seen in a few scattered cells using the PAb240 antibody. The topographical distribution of wild type p53 was not related to proliferating areas but, rather, to short-lived populations of cells. Conclusions-Immunostaining of wild type p53 is demonstrable not only in its nuclear form using antibody PAb240 but also in its common cytoplasmic- perinuclear localisation in normal tissues using the PAb248 monoclonal antibody. This opens up new possibilities for its study in both physiological and pathological conditions.",
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AU - Pezzella, F.

AU - Micklem, K.

AU - Turley, H.

AU - Pulford, K.

AU - Jones, M.

AU - Kocialkowski, S.

AU - Delia, D.

AU - Aiello, A.

AU - Bicknell, R.

AU - Smith, K.

AU - Harris, A. L.

AU - Gatter, K. C.

AU - Mason, D. Y.

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N2 - Aims-To establish whether PAb248 recognises human p53 as weli as murine p53 and if so, to determine its distribution in normal tissues. Methods-The ability of PAb248 to recognise human p53 was established by analysis of the human osteosarcoma derived Saos-2 celil line, which lacks the p53 gene, before and after transfection with p53 cDNA, using western blotting and immunoprecipitation. Immunostaining on normal tissues and cell lines was carried out using an immunoperoxidase technique. The two anti-p53 antibodies PAb 240 and DO-7 were used as controls. Results-The anti-p53 PAb248 monoclonal antibody stained the Saos-2 cell line after, but not before, transfection with p53 cDNA. Both western blots and immunoprecipitations performed with this antibody revealed a 53 000 molecular weight band. With immunostaining, this antibody detects p53 protein in most lymphoid and human epithelial cells in a cytoplasmic-perinuclear localisation that has not been described before. In the same tissues nuclear staining could be seen in a few scattered cells using the PAb240 antibody. The topographical distribution of wild type p53 was not related to proliferating areas but, rather, to short-lived populations of cells. Conclusions-Immunostaining of wild type p53 is demonstrable not only in its nuclear form using antibody PAb240 but also in its common cytoplasmic- perinuclear localisation in normal tissues using the PAb248 monoclonal antibody. This opens up new possibilities for its study in both physiological and pathological conditions.

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