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 language | English |
|---|---|
| Pages (from-to) | 592-596 |
| Number of pages | 5 |
| Journal | Journal of Clinical Pathology |
| Volume | 47 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - 1994 |
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ASJC Scopus subject areas
- Health Policy
- Pathology and Forensic Medicine
Cite this
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 journal › Article
}
TY - JOUR
T1 - Antibody for detecting p53 protein by immunohistochemistry in normal tissues
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.
PY - 1994
Y1 - 1994
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.
AB - 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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UR - http://www.scopus.com/inward/citedby.url?scp=0028641567&partnerID=8YFLogxK
U2 - 10.1136/jcp.47.7.592
DO - 10.1136/jcp.47.7.592
M3 - Article
C2 - 8089212
AN - SCOPUS:0028641567
VL - 47
SP - 592
EP - 596
JO - Journal of Clinical Pathology - Clinical Molecular Pathology
JF - Journal of Clinical Pathology - Clinical Molecular Pathology
SN - 0021-9746
IS - 7
ER -