Promega, N2721, HaloTag® Ubiquitin Proteasome System Fusion Vectors

Regulation of cellular protein homeostasis is critical for maintaining cell health and often altered by cellular treatments or disease states. The majority of proteins and their abundance are regulated via the ubiquitin proteasome system (UPS), which uses ubiquitin conjugation to signal proteins that should be trafficked to the proteasome for degradation. An emerging modality for small-molecule drug development is targeting proteins for degradation via the UPS with small-molecule degraders such as PROTACs or molecular glues.
The HaloTag® fusion vectors are designed to be used as the fluorescent acceptor in NanoBRET® assays that monitor interactions of a specific target protein along the UPS pathway. NanoBRET® technology is a proximity-based method dependent upon energy transfer from a luminescent donor target protein to a fluorescent acceptor. In these assays, the target protein fused to NanoLuc® luciferase or a subunit of NanoBiT® enzyme and serves as the bioluminescent BRET energy donor. The HaloTag® fusions are labeled with the fluorescent HaloTag® NanoBRET® 618 Ligand and serve as the energy acceptor. When the donor and acceptor proteins interact, energy transfer occurs, resulting in increased NanoBRET® signal.