Gamma Secretase Activity Assay Cell Line

Reference: P30702

Gamma Secretase Activity Assay Cell Line

Gamma Secretase Activity Assay Cell Line is an Alzheimer’s Disease in vitro model to identify γ-secretase activity inhibitors. It consists in U2OS cell line stably expressing green fluorescent APP-C99 construct, which contains the γ-secretase cleaving site. To perform the assay, cells are incubated with test compounds during 24 hours to evaluate their effects on APP Processing. After incubation, nucleus could be stained with DAPI and retained fluorescent APP vesicles are detected by fluorescence using image analysis algorithms.

Innoprot offers this assay as a “stable cell line” in cryopreserved vials. Each vial of U2OS cell line stably expressing human tGFP-APP-C99 contains 3 million cells and Innoprot provides 2 vials of cells with each order. It is guaranteed that these cells will expand in more than 30 passages following the cell culture manual.

9.995,00

One of the most critical pathological features of Alzheimer’s disease (AD) is the accumulation of β-amyloid (Aβ) peptides that form extracellular senile plaques in the brain. Increasing evidence suggests that small soluble oligomers of Aβ are the toxic form of the peptide and may instigate AD. The Aβ peptide is the amyloid precursor protein (APP) digestion product. β-secretase and γ-secretase proteolysis releases Aβ from the cell. In addition to APP, γ-secretase also cleaves many other type I transmembrane (TM) protein substrates. As a crucial enzyme for Aβ production, γ-secretase is an appealing therapeutic target for AD.Gamma Secretase Activity Assay Cell Line from Innoprot allows the identification of γ-secretase activity inhibitors.

Innoprot performs High Throughput Imaging assays using this model to identify γ-secretase activity inhibitors. Results indicates that the pharmacological inhibition of γ-secretase implicated in AD remains a valid strategy for drug screening. After treatments, we obtain images and analyze them using Attovision software from Becton Dickinson and we quantify fluorescent vesicles into the cytoplasm.

Results indicate that the detecting dynamyc range is dependent on the inhibitor biophysics and biochemical characteristics and the treatment time. This retention assay shows an average of Z´= 0.79+/- 0.02 for High Content Screening with a 24 hours treatment.

 

Applications

  • This model permits evaluate lybraries of compounds, candidates to γ-secretase inhibitors, in living cells studying the vesicles retention
  • Gamma Secretase Activity Assay provides a strategy to evaluate drug against γ-secretase activity without the necessity to be permeable.
  • This model allows to analyse in the space and time the compund effect in a multiparametric manner.
  • Reference: P30702
  • Size/Quantity: 2 vials containing 3x106 cells / vial
  • Product Use: For research use only
  • Shipping Conditions: Dry Ice

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