Amylase Fluorometric (Green) Assay Kit, EnzChek® from Molecular Probes (Invitrogen)

The EnzChek Amylase Assay Kit (E-11954) provides the speed, high sensitivity and convenience required for measuring a-amylase activity or for screening inhibitors in a high-throughput format. The EnzChek kit contains a starch derivative that is labeled with the BODIPY FL dye to such a degree that the fluorescence is quenched. The DQ starch substrate is efficiently degraded by a-amylase to yield highly fluorescent fragments. The accompanying increase in fluorescence is proportional to amylase activity and can be monitored with a fluorescence microplate reader, minifluorometer or standard fluorometer.

α-Amylase purified from Bacillus sp. is provided with the EnzChek Amylase Assay Kit to serve as a control enzyme. Using 12.5 g/mL of the DQ starch substrate and a 60 minute incubation period at room temperature, the assay can detect the activity of this enzyme down to a final concentration of 1 10-4 U/mL (0.3 g protein/mL), where one unit is defined as the amount of enzyme required to liberate 1 mg of maltose from starch in 3 minutes at 20C, pH 6.9. At high enzyme concentrations (e.g. 2 mU/mL and higher), incubation times can be as short as 5 minutes.