β-Catenin/Tcf Protein-Protein Interaction FP Assay
For Investigation of the Cancer-Related Wnt-Pathway
Schematic Overview of the Fluorescence
- Echelon’s β-catenin/Tcf protein-protein interaction assay is a fluorescence polarization (FP) assay designed for high throughput screening in a 384-well format for small molecule inhibition or the β-catenin:Tcf protein-protein interaction.
- Echelon’s β-catenin/Tcf protein-protein interaction assay is a fluorescence polarization (FP) assay designed for high throughput screening in a 384-well format for small molecule inhibition or the β -catenin:Tcf protein-protein interaction. The β-catenin protein used in this assay is a partial protein which includes the domain that binds to the human Tcf. A selected short peptide sequence of the human Tcf with TAMRA label (Tcf-TAMRA) is used in the assay. When β-catenin binds to the Tcf-TAMRA, the TAMRA fluorophore’s rotational movement becomes slower due to the increase in molecular mass and thus the emitted light remains polarized (see figure on the left). Therefore, the inhibition between β-catenin and Tcf-TAMRA binding can be monitored by the change in polarization of light. PKF118-310 (Lepourcelet 2004) is included in the assay as a control inhibitor.
- Lepourcelet M, Chen YN, France DS, Wang H, Crews P, Petersen F, Bruseo C, Wood AW, Shivdasani RA: Small-molecule antagonists of the oncogenic Tcf/beta-catenin protein complex. Cancer Cell. 2004 Jan;5(1):91-102. Pubmed
- Zhang M, Huang Z, Yu B, Ji H: New homogeneous high-throughput assays for inhibitors of ß-catenin/Tcf protein-protein interactions. Anal Biochem. 2012 May 1;424(1):57-63. Pubmed