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Assay Method Information

Assay Name:  Inhibition Assay
Description:  In this study, the inhibition assay of yeast enzyme was performed in 100 mM phosphate buffer pH 7.0 at 25C with minor changes, according to the methods reported previously. [Saijyo et al., J. Oleo. Sci., 57:431-435] To obtain IC50 value of the inhibitors, a reaction mixture of yeast α-Gls (0.025 U/mL), 0.1 mM pNPG and increasing concentration of each inhibitor was used. Also the inhibition mode was determined for those inhibitors exhibited promising inhibitory activity (C1-C3) by incubating the yeast enzyme solution with the increasing concentration of pNPG (0.1-2 mM), in the presence of different concentration of each inhibitor. Moreover, to explore role of the synthetic compounds on inhibition of mammalian enzyme, mouse α-Gls was extracted and prepared in 100 mM potassium phosphate buffer (pH 7.0), containing 5 mM EDTA. [Zhang et al., Food Chem., 126:466-471] After centrifugation, the supernatant was dialyzed against 10 mM potassium phosphate buffer (pH 7.0) containing 0.
Affinity data for this assay
 

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Last update November 1, 2007
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