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


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タイトル: Facile assay of telomerase activity utilizing a DNA-detectable chemiluminogenic reagent.
著者: Tonooka, Keiko / Kabashima, Tsutomu / Shibata, Takayuki / Tang, Chenhong / Yu, Zhiqiang / Kai, Masaaki
発行日: 2008年 4月
出版者: The Japan Society of Analytical Chemistry
引用: Analytical sciences, 24(4), pp.471-475; 2008
抄録: 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
権利: Copyright (c) 2008 by The Japan Society for Analytical Chemistry
資料タイプ: Journal Article
原稿種類: publisher
出現コレクション:050 学術雑誌論文

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

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