LifeSensors’ UbiTest is a medium-throughput platform for measuring the polyubiquitylation levels of protein(s) of interest (POI). UbiTest utilizes TUBE technology to enrich polyubiquitylated proteins. This fraction is analyzed by immunoblotting techniques with and without pan-selective DUB digestion. The appearance or increase in the intensity of the band corresponding to the POI indicates that the POI is polyubiquitylated in the studied condition. Identification of the type of ubiquitin linkage can provide insights into the cellular functions of E3s and substrates.
- Cell-based assays to measure endogenous substrate ubiquitination.
- Conversion of polyubiquitin smear of the POI to a single band in the DUB-treated fraction simplifies the quantification.
- Overcome the issue of target-specific primary antibody failing to detect ubiquitinated target protein due to epitope masking by the bulky ubiquitin chain.
- Identify the polyubiquitin linkages on the POI by digesting with linkage-specific DUBs.
- Study the effect of PROTACs or small molecules.