Wnt-Frizzled Pathway

Wnt signaling plays a pivotal role in development and is implicated in a variety of disease states ranging from cancer, stem cell loss to kidney and reproductive tract defects. Wnt ligands are associated with both β-catenin dependent (canonical) pathway and β-catenin independent pathway, of which the canonical pathway has been extensively researched. DiscoveRx offers a novel cell-based, chemiluminescent assay that measures effects of compounds on any of the upstream targets that effect β-catenin translocation.


—Features and Benefits

  • Novel cell-based assay that measures downstream β-catenin translocation
  • Whole cell, chemiluminescent assay
  • Avoid lengthy reporter gene assays

—Applications

  • Assay enables the study of:  frizzled receptor activation, upstream GSK 3β inhibition
  • Detect small molecules, siRNA or functional antibodies

—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 42 AA enzyme fragment, ProLabel® is appended to the protein of interest (β‐catenin). The larger enzyme fragment EA (Enzyme Acceptor) resides in the nucleus. Activation of the Wnt signaling pathway initiates β‐catenin translocation to the nucleus that forces complementation of the two β‐Gal enzyme fragments. This action results in the formation of fully complemented β‐galactosidase enzyme, the activity of which is measured using chemiluminescent substrate. The signal can be read on any standard luminometer.

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