HaloPROTAC3 is a small-molecule degrader that specifically binds to and degrades HaloTag and its fusion partners in live cells and can be used to study the degradation of targets that would be challenging with target-specific PROTACs. HaloPROTAC3 binds irreversibly to HaloTag and HaloTag® target fusions, recruiting them via co-engagement with an E3 ligase component (VHL) to active E2/E3 ubiquitin ligase complexes and leading to ubiquitination and subsequent degradation by the proteasome.
The ent-HaloPROTAC3 is the enantiomeric compound of HaloPROTAC3 and serves as a negative control. ent-HaloPROTAC3 has the same molecular weight and general structure as HaloPROTAC3 but contains d-hydroxyproline and d-valine residue modifications allowing it to bind HaloTag but not VHL. Use ent-HaloPROTAC3 to confirm that degradation of the HaloTag® fusion protein is mediated through VHL engagement and the PROTAC mechanism.
We also provide a separate NanoLuc®-HaloTag® fusion vector, NanoBRET™ Positive Control (Cat.# N1581), that can be used for transient transfection expression and degradation with HaloPROTAC3. See the technical manual for an optional protocol using the luminescence functionality of the NanoBRET™ Positive Control to quantify degradation without the use of antibodies.