Human BRD4/CRBN PROTAC Binding Kit (test)

Detection Principle:

The kit utilizes homogeneous time-resolved fluorescence technology (TR-FRET) to determine the interaction between the DDB1/CRBN complex and BRD4 mediated by the molecular glue BET Degrader-1 and the test compound. BRD4 contains bromodomain 1 and 2. This method enables a simple and rapid high-throughput detection of small molecules that can mediate the interaction between the DDB1/CRBN complex and BRD4.

As shown in the figure, the interaction between DDB1/CRBN and BRD4 is detected using Eu-labeled anti-Tag1 antibody (TR-FRET donor) and Accepter-labeled anti-Tag2 antibody (TR-FRET acceptor). Since the molecular glue BET Degrader-1 mediates the interaction between DDB1/CRBN and BRD4, the donor antibody and the acceptor antibody are in close proximity. The excitation of the donor antibody triggers fluorescence resonance energy transfer (FRET) to the acceptor antibody, causing the acceptor antibody to specifically emit a signal with a wavelength of 665nm. This specific signal is proportional to the degree of interaction between BET Degrader-1 and DDB1/CRBN, as well as BRD4. This homogeneous experiment is easy to operate and requires no washing steps.