PROteolysis TArgeting Chimeras (PROTACs) have emerged as novel therapeutics to target the undruggable proteome. PROTACs are heterobifunctional small molecules that both simultaneously bind to a target protein and a ubiquitin (Ub) E3 ligase. This subsequently leads to ubiquitination and degradation of the target protein.
LifeSensors has developed a high-throughput plate-based method to directly monitor PROTAC-mediated or E3 mediated ubiquitination and degradation of target proteins. This method is rapid, efficient, and quantitative and works with endogenous proteins and cell lines. There is no need for stable cell line production or tagging your target protein and creating artifacts. In other words, you can create native conditions to gather the most accurate data possible.
For evaluating PROTACs In vitro, we recommend using our PA770: PROTAC® In Vitro Ubiquitination Assay Kit. This kit is specially designed to measure the performance of PROTACs In vitro, and is available off the shelf for Cereblon, VHL, and MDM2. Other E3 ligases can be optimized for this format as a custom order upon request.
For evaluating molecular glues, we recommend using our MG780: Molecular Glue In Vitro Ubiquitination Assay Kit. This kit is specially designed to measure the performance of molecular glues In vitro.
PROTAC Assay Kit Components:
| Coated plate: | One pre-coated and blocked 96-well strip PROTAC assay plate (Store at -80°C) |
| Blocking concentrate (BC): | 5mL of 5X blocking agent for antibody dilution (Store at 4°C) |
| Plastic plate seals: | Two plate seals |
| Detection reagent 1 (DR1): | 1.0mL vial of DR1 reagent (Store at 4°C) |
| Detection reagent 2 (DR2): | 1.0mL vial of DR2 reagent (Store at 4°C) |
| MG132: | 25µL vial (10 mM in DMSO; Store at -20°C) |
| PR-619: | 25µL vial (22 mM in DMSO; Store at -20°C) |
| 1,10-phenanthroline: | 100µL vial (500 mM in DMSO; Store at -20°C) |