The E3LITE Customizable Ubiquitin Ligase Kit is a robust assay that allows for the quantification of the E3 ubiquitin ligase activity. The kit uses a plate that is pre-coated with a proprietary polyubiquitin capture reagent. The resulting product of the E3 ligase-mediated in vitro ubiquitin reaction is measured and represents a direct measure of the E3 ligase activity. The E3LITE Customizable Ubiquitin Ligase Kit represents a singular platform for the focused investigation of any E2/E3 enzyme pair, as well as any particular polyubiquitin linkage type. This assay has been validated for high throughput screening (HTS) and has been employed in the discovery of novel compounds that selectively modulate E3 ligase function.
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- Ubiquitin E1 Activating Enzyme
- Choice of Ubiquitin E2 Conjugating Enzyme(s)
- E1-E2-E3 Control
- Choice of Recombinant Ubiquitin
- Detection Reagent 1
- Linear Polyubiquitin Control
- (2) modular 96- or 384-well pre-coated plates
|Storage||All components should be stored at -80°C.|
|Stability||All components, except for the pre-coated plates, are stable for at least 2 months when stored at -80°C.
Plates must be used within 30 days.
The selection of appropriate streptavidin reagent is critical to generating a robust signal. We do not recommend using avidin-horseradish peroxidase (HRP) conjugates.