Background
The E2scan™ Kit version 2 is designed to facilitate the identification of E2 conjugating enzymes which support the ubiquitylation of substrate proteins catalysed by an E3 ligase and/or the auto-ubiquitylation of the E3 ligase itself. The kit contains an E2scan™ plate: A panel of 34 E2 conjugating enzymes arrayed in duplicate across a 96-well plate plus all the components (E1 enzyme, control E3 ligase, ubiquitin, ATP, assay buffer) required to perform a complete substrate ubiquitylation or auto-ubiquitylation assay using your E3 ligase and/or E3 ligase/substrate combination of choice. Through the addition of a substrate mix (E1, E3, substrate, and ubiquitin) and then ATP to the wells of an E2scan™ plate you can screen the E2 conjugating enzyme panel in just one hour. Reaction products may be analysed as required, for example by SDS-PAGE or Western blotting using specific antibodies.
Utility of the E2scan™ Kit version 2
Ubiquigent's E2scan™ Kit version 2 will help you to address many fundamental questions in your ubiquitin system research. Potential kit applications are listed below: - Identify the E2(s) which couple with your E3 ligase of interest.
- Determine which E2s support auto and/or substrate-ubiquitylation with your E3.
- Help define mechanisms of E3/E2 interactions through the comparison of E2scan™ data between wild-type and modified versions of the E3 under evaluation.
- Investigate E3 intra-family E2 selectivity.
- Explore the E2 ‘priming theory'.
- Explore potential E2 inhibitors and/or activators.