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Two-enzyme coupled fluorometric assay of urinary dipeptidase
Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
1995, v.8 no.3, pp.359-364
Park,
H. S., &
We,
J.
S.
(1995). Two-enzyme coupled fluorometric assay of urinary dipeptidase. Analytical Science and Technology, 8(3), 359-364.
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Abstract
Urinary dipeptidase(Udpase) was assayed by fluorometric analysis of NADH which was produced from an indicator enzyme, L-alanine dehydrogenase. The reaction mixture was consisted of a dipeptide(L-ala-L-ala), <TEX>${\beta}-NAD^+$</TEX>, L-alanine dehydrogenase in 12.5 mM sodium carbonate buffer, pH 9.0, and urinary dipeptidase which initiated the reaction. The fluorescence intensity of NADH was measured as a function of time with the excitation wavelength at 340nm and emission at 460nm. Comparison of this fluorometric method with the conventional spectrophotometric method utilizing glycyldehydrophenylalanine(Gdp) as substrate provided the correlation coefficient of 0.996 and increased the sensitivity more than ten times.
- keywords
- urinary dipeptidase(Udpase), fluorometric assay, L-alanine dehydrogenase, glycyldehydrophenylalanine (Gdp)
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