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dc.contributor.authorAli, A
dc.contributor.authorChopra, Rajesh
dc.contributor.authorRobertson, J
dc.contributor.authorTesta, Nydia G
dc.date.accessioned2009-11-13T15:21:59Z
dc.date.available2009-11-13T15:21:59Z
dc.date.issued2000-12
dc.identifier.citationDetection of hTERT protein by flow cytometry. 2000, 14 (12):2176-81 Leukemiaen
dc.identifier.issn0887-6924
dc.identifier.pmid11187908
dc.identifier.urihttp://hdl.handle.net/10541/86187
dc.description.abstractTelomerase 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.
dc.language.isoenen
dc.subjectLeukaemiaen
dc.subject.meshCell Differentiation
dc.subject.meshDNA-Binding Proteins
dc.subject.meshFlow Cytometry
dc.subject.meshHL-60 Cells
dc.subject.meshHumans
dc.subject.meshLeukemia, Lymphocytic, Chronic, B-Cell
dc.subject.meshRNA
dc.subject.meshTelomerase
dc.titleDetection of hTERT protein by flow cytometry.en
dc.typeArticleen
dc.contributor.departmentDepartment of Experimental Haematology, Paterson Institute for Cancer Research, Manchester, UK.en
dc.identifier.journalLeukemiaen
html.description.abstractTelomerase 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.


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