| Assay Method Information | |
| | Biochemical Assay |
| Description: | The assay based on fluorescenceresonance energy transfer was carried out with BACE1 enzyme at pH 4.5 with a substrate, H-Lys(DABSYL)-SEVNLDAEFR-Gin-(LY) (Merck) according to Ermolieff's approach15. Briefly, the assay was carried out by incubation of the substrate peptide (5 μM), BACE1 enzyme (550 ng/mL) that was expressed and purified from insect expression system and varied concentrations of compounds at pH 4.5. After 1 h incubation at 37°C, stop buffer (3 M sodium acetate) was introduced into the reaction mixture to stop the reaction. Finally, the fluorescence intensity was measured on a TECAN GENios reader (Tecan) at room temperature with excitation at 420 nm and emission at 530 nm. |
| Affinity data for this assay | |
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