| Assay Method Information | |
| | PHGDH Enzyme Assay |
| Description: | To evaluate the effects of compounds on PHGDH activity, compounds were first pre-incubated with enzyme samples in the assay buffer (25 mM HEPES, pH 7.1, 400 mM KCl, 5 μM phosphopyridoxa (PLP), 0.5 mM α-ketoglutarate, 150 μM NADH, 1 mM DTT) for 10 min at 25℃, then the reaction was started by adding L-phospho-O-serine (Pser). Each compound was dissolved in DMSO at a final concentration of 5%,which did not affect the assay signal. Fluorescence signals were recorded for 3 min with a kinetics mode program using on a plate reader (Synergy, Biotek). |
| Affinity data for this assay | |
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