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Facile assay of telomerase activity utilizing a DNA-detectable chemiluminogenic reagent.


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Title: Facile assay of telomerase activity utilizing a DNA-detectable chemiluminogenic reagent.
Authors: Tonooka, Keiko / Kabashima, Tsutomu / Shibata, Takayuki / Tang, Chenhong / Yu, Zhiqiang / Kai, Masaaki
Issue Date: Apr-2008
Publisher: The Japan Society of Analytical Chemistry
Citation: Analytical sciences, 24(4), pp.471-475; 2008
Abstract: Telomerase shows increased activity in most human cancers and germ line cells, but not in normal human somatic cells. We describe a novel chemiluminescence method for the facile assay of telomerase activity in human cells. The telomerase substrate was incubated with the cell lysate containing various amounts of telomerase, and then the telomerase product was amplified by the polymerase-chained reaction (PCR). The PCR products were separated from the excess substrate, primer and deoxyribonucleotide triphosphates by a centrifugal filter, which distinguished different molecular sizes. The isolated products were reacted with a DNA-detectable chemiluminogenic reagent, 3,4,5-trimethoxyphenylglyoxal. The proposed assay method gave linearity for the telomerase activity in 100 to 10000 cells (r2 = 0.997), and allowed the assay not only of lower activity, but also of higher activity of telomerase without the requirement of any special labeled-PCR primers in the assay system.
URI: http://hdl.handle.net/10069/18702
ISSN: 09106340
DOI: 10.2116/analsci.24.471
PubMed ID: 18403837
Rights: Copyright (c) 2008 by The Japan Society for Analytical Chemistry
Type: Journal Article
Text Version: publisher
Appears in Collections:Articles in academic journal

Citable URI : http://hdl.handle.net/10069/18702

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