PI3 Pathway

PI3-K/AKT signaling regulates cellular processes such as proliferation, growth, survival, apoptosis, and the up regulation of hypoxia-related proteins. Activation of this pathway involves various receptor tyrosine kinases, such as IGFR1 receptor for insulin. DiscoveRx offers novel, non-imaging , cell-based and biochemical assays to study various aspects of the PI3 Kinase Pathway. Ready to screen, biosensor cell lines allows you to study receptor activation, phosphorylation or downstream FOXO translocation in a HTS-friendly cell-based format. Our universal, non-antibody ADP accumulation assay provides a screening-friendly biochemical assay format, for identifying PI3 or AKT inhibitors.

PathHunter® U2OS FOXO3 Nuclear Translocation Assay


—Features and Benefits

  • Novel cell-based assay that measures downstream FOXO translocation
  • Whole cell, chemiluminescent assay
  • Single addition, HTS-friendly format

—Applications

  • Screen for inhibitors to:  PI3 Kinase and AKT
  • Study molecules affecting FOXO3 nuclear translocation

Assay Principle

The PathHunter assay monitors the interaction of two proteins in a whole cell, homogeneous assay format using enzyme fragment complementation. The cells have been engineered to express two complementing fragments of β-Gal within different cellular compartments. In this system, a small enzyme fragment, ProLink is attached to the protein of interest, FOXO-3. The larger enzyme fragment EA (Enzyme Acceptor) resides in the nucleus. Activation of the pathway initiates FOXO-3 translocation to the nucleus that forces complementation of the two enzyme fragments. This action results in the formation of fully complemented β-galactosidase enzyme, the activity of which is measured using chemiluminescent PathHunter Detection Reagents. The assay can be used to study AKT and PI3 Kinase inhibitors.View Categories: Pathway Assays

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