Detection of human topoisomerase II alpha in cell lines and tissues: characterization of five novel monoclonal antibodies.
J Histochem Cytochem
; 45(2): 251-63, 1997 Feb.
Article
em En
| MEDLINE
| ID: mdl-9016314
ABSTRACT
We report five novel monoclonal antibodies (Ki-S1, Ki-S4, Ki-S6, Ki-S7, and Ki-S8) reactive with a proliferation-related nuclear antigen. In immunoprecipitation and Western blot experiments using crude nuclear extracts, they recognized a protein of 170 kD that, after proteolytic digestion of the immunoprecipitate and sequencing of the resulting peptides, was identified as the alpha-isoform of human topoisomerase II. This was confirmed by testing the antibodies on a highly purified enzyme preparation. Crossreactivity with topoisomerase II beta was ruled out by testing the antibodies on crude extracts from yeast cells expressing the beta-isoform exclusively. The antibodies bind the antigen with different affinities and at different epitopes, apparently located within the carboxyl third of the enzyme. All five antibodies are suitable for archival material after adequate antigen retrieval, thereby enabling retrospective studies. This report illustrates the tissue and subcellular distribution of the antigen through the cell cycle by immunohistochemistry and confocal fluorescence microscopy. The antibodies will be useful tools in further analysis of morphological and functional aspects of topoisomerase II and may serve diagnostic purposes, as well as providing prognostic information in tumor pathology.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Topoisomerases Tipo II
/
Isoenzimas
/
Anticorpos Monoclonais
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Antígenos de Neoplasias
Tipo de estudo:
Diagnostic_studies
/
Observational_studies
Limite:
Humans
Idioma:
En
Revista:
J Histochem Cytochem
Ano de publicação:
1997
Tipo de documento:
Article