| Assay Method Information | |
| | AlphaScreen Assay |
| Description: | BACE1 activity can be assessed in an AlphaScreen assay. Compounds to be assessed (e.g. compounds as described in the above examples) are serially diluted 1:4 in DMSO, then plated by diluting 10 μL of compound with 40 μl of assay buffer (1×PBS, 0.01% Tween-20, pH 7.0, 0.05% BSA). The diluted compound is transferred (4 μL per well) to a 386 well assay plate and 4 μL of BACE1 enzyme (20 mg/mL) is added and incubated for 15 minutes. This assay uses a probe that is a BACE1 binding sight ligand linked to biotin, which can be displaced by test compound dependent on compound binding affinity. This of biotinylated probe (4 μL) is added and incubated for 1 hour, followed with 4 μL of donor and acceptor bead mixture and incubated for 2 hours. The fluorescent signal is read on a plate reader and the value as a function of compound concentration is used to determine the IC50. |
| Affinity data for this assay | |
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