Does My Target Become Ubiquitinated?
We also offer biochemical methods to monitor ubiquitination of a target protein. The example shown here detects ubiquitination of the E3 ligase Cbl-b using Lumit™ Immunoassay technology.
Schematic overview of a biochemical approach to monitor target protein ubiquitination. In the presence of necessary ubiquitination components E1, E2 and ATP, the Lumit™ Immunoassay can measure the interaction between biotinylated ubiquitin and GST-Cbl-b.
Measuring autoubiquitination of Cbl-b. Panel A. A concentration-dependent increase in Cbl-b-GST ubiquitination in the presence of ATP causes increase in luminescence signal. As the process is ATP-dependent, in the absence of ATP there is a low, background signal. Panel B. A dilution series of unlabeled ubiquitin was used to compete off the biotinylated ubiquitin and results in concentration-dependent decrease in luminescence.