KeyTec® TR-FRET Human cereblon binding kit

KeyTec® TR-FRET human cereblon binding kit is designed for the binding analysis of cereblon ligands/inhibitors to wild-type human cereblon. This kit provides a tag1- cereblon/DNA damage-binding protein 1 complex ( Tag1-CRBN/DDB1 ), a Thalidomide-HX*1 (a cereblon inhibitor), 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 cereblon 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-CRBN/DDB1 and the Thalidomide-HX*1 brings the TR-FRET donor and acceptor into close proximity, enabling resonance energy transfer (RET) upon excitation and generating a TR-FRET signal. Cereblon ligands/inhibitors compete with Thalidomide-HX*1 for binding to the Tag1-CRBN/DDB1. The intensity of TR-FRET signal is inversely proportional to the apparent affinity of Cereblon ligands/inhibitors.

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

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