Method for quantitative analysis for prolyl hydroxylase activity using fluorescence polarization
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United States of America Patent
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Apr 6, 2010
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app pub date -
Dec 6, 2005
filing date -
Jul 22, 2005
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Abstract
The present invention relates to a method for analyzing the interaction between HIF-1 peptide and VBC protein using fluorescence polarization, more precisely, a method for quantitative analysis of formation of HIF-1-VBC protein complex which is composed of the steps of 1) preparing a fluorescent probe by attaching a fluorescein to hydroxyproline containing HIF-1 peptide; 2) reacting the fluorescent probe with VBC protein; and 3) measuring the fluorescence polarization of the above reactant and then comparing the fluorescence polarization with that of the fluorescent probe itself to investigate the changes of fluorescence polarization; a method for screening an inhibitor of the binding of HIF-1 peptide and VBC protein using the above method; and a method for analyzing the activity of prolyl hydroxylase using the above method. The method of the present invention enables simple analysis of the interaction between HIF-1 peptide and VBC protein by observing changes of fluorescence polarization and thus, it can be effectively used for high-speed screening using a well plate.
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