Detection of hTERT protein by flow cytometry

  1. Lookup NU author(s)
  2. Dr Ased Ali
Author(s)Ali AS, Chopra R, Robertson J, Testa NG
Publication type Article
JournalLeukemia
Year2000
Volume14
Issue12
Pages2176-2181
ISSN (print)0887-6924
ISSN (electronic)1476-5551
Full text for this publication is not currently held within this repository. Alternative links are provided below where available.
Telomerase is a telomere-specific DNA polymerase consisting of protein and RNA components, which is activated in germline cells and the majority of cancers and serves to counter the consequences of telomere shortening. The protein component, hTERT, is believed to be the catalytic subunit of human telomerase and its expression at the mRNA level correlates well with telomerase activity in vitro. Current techniques for assaying telomerase activity detect only the mean activity in a sample and are unable to isolate specific cell sub-populations. This report describes the development and validation of a cellular, immunofluorescence-based flow cytometry assay that allows detection of intranuclear hTERT while maintaining identifiable cell population characteristics. The assay was shown to be both sensitive to changes in telomerase expression and was semi-quantitative. In both cell line differentiation experiments and in primary cells, a good correlation existed between hTERT expression measured by flow cytometry and telomerase activity detected by the telomeric repeat amplification protocol (TRAP). The method developed offers a quick, simple and reproducible cellular-based assay for hTERT expression. This assay will provide a useful, new tool for future investigations, facilitating the analysis of hTERT expression in mixed cell populations.
PublisherNature Publishing Group
URLhttp://dx.doi.org/10.1038/sj.leu.2401950
DOI10.1038/sj.leu.2401950
Actions    Link to this publication
Share