| Assay Method Information | |
| | PSMA inhibitory Activity Assay |
| Description: | This test example is used to display the results of the PSMA inhibitory activity tested for each compound and comparative compound. A LNCaP cell lysate (total protein concentration: 125 μg/mL) was prepared in advance. 25 μL of the cell lysate, 25 μL of the inhibitor and 25 μL of N-acetylaspartylglutamate (NAAG, 16 μM) were incubated together in a 96-well plate (Costar Assay Plate, Cat#. 3925) at 37° C. for 180 min. The amount of glutamate released by hydrolysis of NAAG was measured using an Amplex Red Glutamic Acid Kit (Molecular Probes Inc., Eugene, OR) working solution (50 μL) after incubation for 30 min Fluorescence were measured using Synergy H1 Hybrid Reader (BioTek Instruments, Inc., Winooski, Vermont) at excitation wavelength of 530 nm and emission wavelength of 590 nm. An inhibition curve was plotted by a semi-logarithmic method. ICso values were calculated as the inhibitor concentration at which the enzyme activity was inhibited to 50%. |
| Affinity data for this assay | |
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