| Assay Method Information | |
| | Arginase Inhibitory Activity Test |
| Description: | Specifically, all reagents required for evaluation were diluted in a reaction buffer (8 mM NAHPO4, 2 mM KH2PO4, pH 7.5, 137 mM NaCl, 2.7 mM KCl, and 0.05% Tween-20), and the compound was dissolved in DMSO and then diluted, and then diluted in the reaction buffer to conform to the desired concentration. 10 μL of the compound and 10 μL of 30 nM Arginase-1 were mixed in a transparent 96-well plate (SPL), and incubated at room temperature for 1 hour or more. After that, 10 μL of 15 mM L-arginine and 3 mM MnCl2 were added to the plate, and then reacted at room temperature for 30 minutes or more. Finally, 30 μL of a 1:1 mixture of reagent A (10 mM o-phthaldialdehyde, 0.4% polyoxyethylene lauryl ether, and 1.8M sulfuric acid in DW) and reagent B (1.3 mM primaquine diphosphate, 0.4% polyoxyethylene lauryl ether, and 3.6 M sulfuric acid in DW) was added to complete the enzyme reaction, and incubated at room temperature for 1 hour or more. The plate after incubation was measured for absorbance at 450 nM using a Flexstation3 multi-mode microplate reader (Molecular Devices). |
| Affinity data for this assay | |
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