Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology

Products

β-Catenin/Tcf Protein-Protein Interaction FP Assay

For Investigation of the Cancer-Related Wnt-Pathway

Schematic Overview of the Fluorescence
Polarization Assay

Description

  • 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.

  • β-catenin is a key mediator for the Wnt pathway. The Wnt pathway is classified as a β-catenin-dependent pathway or β-catenin-independent pathway. The Wnt/β-catenin-dependent pathway is responsible for many cellular regulations such as proliferation, differentiation, and cell-cell communication. Hyperactivated Wnt/β-catenin-dependent pathway triggers progression of many cancers such as colorectal cancers, liver cancer, breast cancer, etc. The transcription of the overactive Wnt/β-catenin-dependent pathway is solely dependent on the nuclear β-catenin:T- cell factor (Tcf) binding. Thus, inhibitors targeting the β-catenin/Tcf protein-protein interaction appear to be a promising therapeutic strategy for cancer. Lately, literature suggests activation of the β -catenin/Tcf signaling by the lipid growth factor, lysophosphatidic acid (LPA), may be involved in neurogenesis as well.

  • 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.

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
K-5900-EC B-Catenin/Tcf Protein-Protein Interaction FP Assay 1 kit 500,00 PDF
  • All prices are in EURO excl. VAT and shipping. Only available in Europe except for France and Italy.

Literature

  • 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