Monoclonal antibodies to human DNA topoisomerase I and the two isoforms of DNA topoisomerase II: 170- and 180-kDa isozymes

Claudia Negri, Roberto Chiesa, Antonella Cerino, Marco Bestagno, Cinzia Sala, Nicoletta Zini, Nadir M. Maraldi, Giulia C B Astaldi Ricotti

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Abstract

Several monoclonal antibodies of different isotypes specific to human DNA topoisomerase I, to 170- and 180-kDa DNA topoisomerase II isozymes, were produced and characterized. The specificity of monoclonal antibodies was confirmed by comparison with polyclonal antibodies by Western blot, by immunoprecipitation of enzyme activity, and by immunoprecipitation of DNA topoisomerases with characterized polyclonal antisera. Morphological studies performed by immunofluorescence indicate that the three groups of monoclonal antibodies (MoAbs) stain the nucleus with characteristic patterns, which can be compared with those obtained with polyclonal antibodies. In particular the MoAbs to the 100-kDa DNA topoisomerase I stain the nucleolus and the nucleoplasm; the MoAbs to 170- and 180-kDa DNA topoisomerase II give completely distinct intranuclear patterns: those to the 170-kDa protein stain mainly the nucleoplasm, whereas those to the 180-kDa protein stain only the nucleolus. The two DNA topoisomerase II isozymes clearly exhibit fluctuations in their expression during cell growth: the 170-kDa isozyme is more abundant during the logarithmic phase of growth, while the 180-kDa isozyme is mainly present during the plateau phase of growth.

Original languageEnglish
Pages (from-to)452-459
Number of pages8
JournalExperimental Cell Research
Volume200
Issue number2
DOIs
Publication statusPublished - 1992

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Type II DNA Topoisomerase
Isoenzymes
Protein Isoforms
Monoclonal Antibodies
Coloring Agents
Immunoprecipitation
DNA Topoisomerases
Growth
Type I DNA Topoisomerase
Antibodies
Fluorescent Antibody Technique
Immune Sera
Proteins
Western Blotting
human TOP1 protein
Enzymes

ASJC Scopus subject areas

  • Cell Biology

Cite this

Monoclonal antibodies to human DNA topoisomerase I and the two isoforms of DNA topoisomerase II : 170- and 180-kDa isozymes. / Negri, Claudia; Chiesa, Roberto; Cerino, Antonella; Bestagno, Marco; Sala, Cinzia; Zini, Nicoletta; Maraldi, Nadir M.; Astaldi Ricotti, Giulia C B.

In: Experimental Cell Research, Vol. 200, No. 2, 1992, p. 452-459.

Research output: Contribution to journalArticle

Negri, Claudia ; Chiesa, Roberto ; Cerino, Antonella ; Bestagno, Marco ; Sala, Cinzia ; Zini, Nicoletta ; Maraldi, Nadir M. ; Astaldi Ricotti, Giulia C B. / Monoclonal antibodies to human DNA topoisomerase I and the two isoforms of DNA topoisomerase II : 170- and 180-kDa isozymes. In: Experimental Cell Research. 1992 ; Vol. 200, No. 2. pp. 452-459.
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AU - Bestagno, Marco

AU - Sala, Cinzia

AU - Zini, Nicoletta

AU - Maraldi, Nadir M.

AU - Astaldi Ricotti, Giulia C B

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