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E3LITE Customizable Ubiquitin Ligase Kit

 

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 quantitative measure of E3 ligase activity, with a detection strategy that does not require additional tagging or labeling of either ubiquitin or ligation machinery. Analyzing E3 ligase 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 polyubiquitylation 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 (25) E2 conjugation enzymes, with a selection representing members from each enzyme class. LifeSensors also offers E2 Selection Kits that provides a panel of (4) E2 conjugating enzymes. 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.

 

Applications
  • 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

 

Benefits
  • 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).