These quantitative, plate-based assays are used to determine the most optimal E2 for the E3 of interest. The E2 can determine the ubiquitin chain-linkage type produced and the ability of your E3 to ubiquitylate substrates. Thus, E2 selection is a vital part for optimal E3 ligase activity.
-Polyubiquitin Chain Identification
Different types of ubiquitination linkage patterns can have different functional consequences for the substrate. Knowing the type of chain and branching can give insights into the signaling mechanism and the role of your E3 or substrate of interest. LifeSensors’ E3 ligase assays represent a very effective method for polyubiquitin chain identification by monitoring the activity of the E3 ligase with WT- and Lysine- mutant ubiquitin.
-E3 Screening and Compound Profiling
Small molecule and PROTAC libraries can be screened using LifeSensors’ E3 ligase assays. These assays can also be used to screen for multiple E3 ligases at the same time, thus, establishing compound selectivity.