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Innoprot

Alzheimer`s Disease in vitro Model - APP Processing Assay (Amyloid-beta synthesis in vitro)

Data Sheet


Alzheimer’s Disease (AD) is characterized by brain depositions of the beta amyloid (bA). The bA is the amyloid precursor protein (APP) digestion product, which is released from the cell after b-secretase and  g-secretase proteolysis.

A novel fluorescence-based assay for screening of inhibitors of the APP Processing pathway has been developed and fully validated with reference compounds. This novel assay is based on a MDCK cell line stably expressing green fluorescent APP, which treated with Aβ peptide formation inhibitors such as secretase activy inhibitors, forms fluorescent spots in a dose-response manner.


APP Processing Assay Details


A cell line stably expressing human APP-tGFP is treated with test compounds for 72 hours to evaluate their effects against Aβ peptide  synthesis.  When  APP  processing  is  stopped  by  any  compound  (ej.  secretase  inhibitors),  retained  fluorescent  APP  spots  are  formed.  After  72  hours,  nucleus  are  stained  with  DAPI  and  cells  with  retained APP spots are detected and quantified by fluorescence using image analysis.


Alzheimer's Disease in vitro Model - Applications

     

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