This simple and robust assay allows for quantification of E3 ubiquitin ligase activity. The kit employs a proprietary reagent to capture polyubiquitin chains formed in an E3 ligase dependent manner. Determination of the amount of product formed is a quantitative measure of the E3 ligase activity, and the detection strategy does not require additional tagging or labeling of either ubiquitin or the ligation machinery. Analyzing E3 ligase activity enables enzyme characterization as well as compound screening for modulators of ubiquitylation, an emerging focus in the drug discovery field. 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.*
The E3LITE Customizable Ubiquitin Ligase Kit is flexible by design, essentially providing a singular platform for the focused investigation of any E2/E3 enzyme pair, as well as any particular polyubiquitin linkage type. In support of this flexible platform, LifeSensors offers the widest array of reagents on the market to meet your research needs. The kit itself can be assembled with any one of more than twenty-four E2 conjugation enzymes, a selection representing members from each enzyme class. For more E2 enzymes, assemble a panel of (4) from E2 Selection panel. The recombinant ubiquitin component of this kit can be wild-type, variants that specifically form K48 linkage types, or variants that form linkages only through one of the seven possible surface exposed lysines.
*E3LITE kit is also available in 384-well customized format (cat.no.UC-384; $450)
**Plates must be used within 30 days of receipt.
This product is protected by one or more US or Foreign patents. Please read the Limited Use Label License to learn more. By purchasing this product, the purchaser agrees to comply to these terms.
Price shown is for the one E2 enzyme included in the kit. Additional (4) E2 panel can be purchased for $195. To order more E2s, please inquire about the price. Email email@example.com for more information, including commercial availability.
- 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-well plates coated with LifeSensors proprietary reagent
|Storage||All components should be stored at -80°C.|
|Stability||All components, except for microtiter 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.
- Quantitative Analysis of Ubiquitin E2 Conjugation or E3 Ligase Activity
- Demonstration of novel E2 or E3 Activity
- High Throughput Screening (HTS) of antagonist or agonists of ligase activity
- Monitor enzyme activity during purification
- Monitor E3 activity in solution phase, with the E2 of your choice
- Plate-based format is amenable to high-throughput screening (HTS)
- Detection system provides robust readout for E3 ligase activity
- E3LITE assay utilizes non-radioactive reporter substrates
- E3LITE assay does not require excitation in the UV range (reducing false positive rate)