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Related Experiment Videos

Fen1 expression: a novel marker for cell proliferation.

E Warbrick1, P J Coates, P A Hall

  • 1Department of Biochemistry, University of Dundee, Scotland, U.K. e.warbrick@dundee.ac.uk

The Journal of Pathology
|April 22, 1999
PubMed
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The anti-Fen1 antibody identifies proliferating cells by detecting the Fen1 protein, a nuclear antigen expressed during the cell cycle. Unlike other markers, Fen1 is not induced by DNA damage, making it ideal for proliferation analysis.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Cancer Research

Background:

  • Identifying cell proliferation is crucial for understanding tissue dynamics and disease progression.
  • Antigens associated with the cell cycle offer precise markers for proliferative populations.
  • The role of Fen1 (Flap Endonuclease 1) in DNA repair remains less understood than its role in replication.

Purpose of the Study:

  • To characterize a novel polyclonal antibody (3220) against Fen1 for various applications.
  • To investigate Fen1's expression, localization, and regulation in response to DNA damage.
  • To evaluate Fen1 as a reliable marker for cell proliferation in histological and cytological samples.

Main Methods:

  • Raised a polyclonal antibody (3220) targeting Fen1.

Related Experiment Videos

  • Utilized western blotting to assess Fen1 levels.
  • Employed immunoprecipitation and immunohistochemical analysis (immunostaining) for subcellular localization studies.
  • Examined Fen1 expression and localization in cultured cells and tissue samples, including response to DNA damaging agents.

    • Main Results:

    • Fen1 is a nuclear antigen expressed by cycling cells.
    • Fen1 co-localizes with PCNA and polymerase alpha during the S phase of the cell cycle.
    • Fen1 expression is topologically regulated in vivo and associated with proliferative populations.
    • No significant changes in Fen1 expression levels or patterns were observed following exposure to DNA damaging agents, in vitro or in vivo.

    Conclusions:

    • The anti-Fen1 antibody is a valuable tool for assessing proliferation in histological materials.
    • Fen1 serves as a reliable proliferation marker, as its expression correlates with the growth fraction and is not affected by DNA damage.
    • Fen1's distinct regulation by the cell cycle, independent of DNA damage response, enhances its utility over markers like PCNA.