Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Label-Free and Real-Time Monitoring of Phosphatase Reactions Using a Phosphate-Specific and Fluorescent Probe

  • Lee, Ji-Hoon (Life Sciences Research Division, Korea Institute of Science and Technology) ;
  • Ahn, Hee-Chul (Life Sciences Research Division, Korea Institute of Science and Technology) ;
  • Shin, Dong-Yun (Life Sciences Research Division, Korea Institute of Science and Technology) ;
  • Ahn, Dae-Ro (Life Sciences Research Division, Korea Institute of Science and Technology)
  • Published : 2008.05.20
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Abstract

A phosphate-specific and fluorescent probe was prepared for label-free phosphatase assays based on fluorescence polarization. By using the probe, dephosphorylation reactions of DNA and protein substrates by calf intestinal alkaline phosphatase (CIP) could effectively be monitored in real-time. Since this assay method does not require additional materials such as labeled substrates and phosphospecific antibodies to obtain fluorescence polarization signals, it is simple, cost-effective, and expected to be useful not only for measuring activity of phosphatases but also for high-throughput screening of phosphatase inhibitors.

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