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Assay Method Information

Assay Name:  Inhibition Assay Against Commercial Glycosidases
Description:  In a 96-well plates, 10µL of commercial enzyme solutions without (control) or with 20µL of inhibitor were incubated at 37°C for 5 min. After addition of 20µL of substrate solution, incubations were prolonged for different time periods: 10 min for β-glucosidase (almond), 5 min for α-glucosidase (saccharomyces cerevisiae) and 8 min for β-galactosidase (bovine liver) and α-galactosidase (green coffee beans) and stopped by addition of 150µL of glycine/NaOH buffer (100mM, pH 10.6). The amount of 4-MU formed was determined at 355nm (excitation) and 460nm (emission). For β-glucosidase (almond), the activity was determined with 4-MU-β-D-Glu (1mM) in sodium acetate buffer (100 mM, pH 5.2). For α-glucosidase (saccharomyces cerevisiae), the activity was determined with4-methylumbelliferyl-α-D-glucopyranoside (4-MU-α-D-Glu, 1 mM) in sodium phosphate buffer (100 mM, pH 7.2). For β-galactosidase (bovine liver), the activity was determined with 4-me
Affinity data for this assay
 

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Last update November 1, 2007
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