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Innoprot

AD Model - Gamma-Secretase Activity Assay

Data Sheet


Alzheimer disease (AD) is characterized by brain depositions of the beta amyloid (βA). The βA is the amyloid precursor protein (APP) digestion product. For the generation of Aβ, APP is first cleaved on the N-terminal side of the Aβ sequence by the β-site APP cleaving enzyme-1 (BACE1), a transmembrane aspartyl protease. The resulting 99-amino acid residue C-terminal fragment (C99) is then cleaved by the γ-secretase to yield Aβ and a C-terminal APP intracellular domain (AICD) fragment.

U2OS cells stably expressing a fluorescent construct containing the APP-C99 fragment (γ-secretase cleaving site) are used in this assay to screen inhibitor of the γ-secretase proteolityc process in living cells. Inhibiting γ-secretase activity, fluorescent aggregates will be formed.


Assay Details


Cells will be incubated with the compounds to be tested during 24 hours using DAPT at 10 uM as positive control. Fluorescent aggregates formation of APP-C99 fragment will be quantified using the BD Pathway HCS Reader and Attovision Compartimentalization Software.


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