| Assay Method Information | |
| | Inhibitory Assay |
| Description: | First, 48.5 μl portions of a solution of substrate peptide (Biotin-XSEVNLDAEFRHDSGC-Eu: X=ε-amino-n-capronic acid, Eu=Europium cryptate) were put into respective wells of a 96-well half area microplate (black microplate, Costar). Then, 0.5 μl of a test compound (DMSO (dimethyl sulfoxide) solution) and 1 μl of Recombinant human BACE-1 (R&D systems) were added to each of the wells, and they were reacted for 3 hours at 30° C. The substrate peptide was synthesized by reacting Biotin-XSEVNLDAEFRHDSGC (PEPTIDE INSTITUTE, INC.) with Cryptate TBPCOOH mono SMP (CIS bio international). The final concentration of the substrate peptide was set to 18 nM, and the final concentration of Recombinant human BACE-1 was set to 7.4 nM. The reaction buffer used was a sodium acetate buffer (50 mM sodium acetate, pH 5.0, 0.008% Triton X-100). |
| Affinity data for this assay | |
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