KeyTec® TR-FRET Human VHL binding kit

KeyTec® TR-FRET human von Hippel−Lindau(VHL) binding kit is designed for the binding analysis of VHL ligands/inhibitors to wild-type human VHL. This kit provides a tag1- VHL/Elongin C/Elongin B protein complex ( Tag1-VCB complex ), a VHL ligand-LA*1 (CAS#1948273-02-6), an anti-tag1 antibody labeled with KeyTec® TR-FRET Solar Eu*2 (mAb anti-Tag1 - Solar Eu) and other detection reagents. These components enable the high-throughput screening of VHL ligands/inhibitors.

This assay is based on a competitive immunoassay method using KeyTec® TR-FRET technology, offering a simple, rapid, highly specific and sensitive, as well as reproducible detection process. The principle, outlined in Figure 1:

The binding of the mAb anti-Tag1 - Solar Eu, the Tag1-VCB complex and the VHL ligand-LA*1 brings the TR-FRET donor and acceptor into close proximity, enabling resonance energy transfer (RET) upon excitation and generating a TR-FRET signal. VHL ligands/inhibitors compete with VHL ligand-LA*1 for binding to the Tag1-VCB complex. The intensity of TR-FRET signal is inversely proportional to the apparent affinity of VHL ligands/inhibitors.

*1  KeyTec® TR-FRET LA: TR-FRET Acceptor

*2  KeyTec® TR-FRET Solar Eu: TR-FRET Donor