Facile Assay of Telomerase Activity Utilizing a DNA-detectable Chemiluminogenic Reagent

• TONOOKA Keiko

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

• KABASHIMA Tsutomu

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

• SHIBATA Takayuki

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

• TANG Chenhong

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

• YU Zhiqiang

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

• KAI Masaaki

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

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.

Journal

• Analytical Sciences

  Analytical Sciences 24 (4), 471-475, 2008

  The Japan Society for Analytical Chemistry

Keywords

• Analytical Chemistry

Details

• CRID: 1390282679232910080
• NII Article ID: 10021099521
• NII Book ID: AA10500785
• DOI: 10.2116/analsci.24.471
• ISSN: 13482246
• ISSN: 09106340
• HANDLE: 10069/18702
• NDL BIB ID: 9452076
• Web Site: https://nagasaki-u.repo.nii.ac.jp/records/19369
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I9452076
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.24.471.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.24.471/fulltext.html
• Web Site: http://www.jstage.jst.go.jp/article/analsci/24/4/24_4_471/_pdf
• Text Lang: en
• Data Source:
  • JaLC
  • IRDB
  • NDL
  • Crossref
  • CiNii Articles
• Abstract License Flag: Disallowed