| Assay Method Information | |
| | DDC-MDH-PEPC-Linked assay |
| Description: | To the wells of a 384-well polypropylene plate, 1 µL DMSO (control) or test compounds at indicated concentrations were added, followed by the addition of 24.5 µL enzyme mix [50 mM Tris-HCl, 50 mM NaCl, 0.015% (w/v) BSA, 5 mM MgCl2, 2 mM β-mercaptoethanol, 760 µM NADH, 330 nM PEPC, 100 µM PLP and 284 nM hDDC, pH 8.05], and 24.5 µL substrate mix [50 mM Tris-HCl containing 50 mM NaCl, 0.015% (w/v) BSA, 5 mM MgCl2, 2 mM β-mercaptoethanol, 10 mM PPPA, 0.49 U MDH and 1.5 mM L-DOPA, pH 8.05]. The plate was sealed and incubated for 1 h in 37 °C. The OD was taken at 340 nm. |
| Affinity data for this assay | |
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