| Assay Method Information | |
| | Glucokinase Enzyme Assay |
| Description: | The GK activity was assessed spectrometrically by a coupled reaction with glucose-6-phosphate dehydrogenase (G6PDH). Briefly, GK catalyzes glucose phosphorylation to generate glucose-6-P, which was oxidized by the G6PDH with the concomitant reduction of NADP. The product NADPH was then monitored by the increase rate of absorbance at 340 nm in a plate reader (SpectraMax 190; Molecular Devices, USA). For EC50 determination, different concentrations of compounds were tested in the assay, and the fold changes in activity versus controls were fitted to sigmoidal curve using a four parameter logistic model in GraphPad Prism 4. |
| Affinity data for this assay | |
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