| Assay Method Information | |
| | Glycogen Phosphorylase Activity Assay |
| Description: | The activity of recombinant human liver GPa in the forward direction was measured by monitoring the production of NADPH. Enzyme activity was assayed at pH 6.8 in phosphate buffer containing beta-NADP, alpha -glucose 1,6-bisphosphate, glucose-6-phophate dehydrogenase, phosphoglucomutase, glycogen, and glucose. The basal rate of hLGPa enzyme activity in the absence of inhibitors (Control) was determined by adding DMSO alone, and a fully-inhibited rate of hLGPa enzyme activity was obtained by adding the positive control test substance, 200 mM caffeine. The reaction was started by adding hLGPa solution (beta-glycerophosphate and cystein, pH 6.8). The reaction took place at room temperature, and the conversion of oxidized beta-NADP to reduced beta-NADPH was measured at 340 nm for 2 h. The hLGPa inhibition IC50 values were calculated using the logistic regression method and SAS software. |
| Affinity data for this assay | |
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