UC101: E3LITE Customizable Ubiquitin Ligase Kit


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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.

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.

Kit Components

  •  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.

Additional information

Plate Size

96-well plate, 384-well plate

E2 Conjugating Enzyme

UBE2D3 (UbcH5c), UBE2L3 (UbcH7), UBE2K (UbcH1), Ubc13, Mms2 (yeast), UBE2D2 (UbcH5b, E2-17K2), UBE2R1 (CDC34), UBE2R2 (Ubc3B), Non-tagged UBE2D1, His6-SUMO tagged UBE2D1, UBE2E2 (UbcH8), UBE2E3 (UbcH9), His6-tagged UBE2H (E2-20K), His6-SUMO tagged UBE2H (E2-20K), UBE2F (NCE2), UBE2M (Ubc12), UBE2N (Ubc13), Non-tagged UBE2T (PIG50), His6-SUMO tagged UBE2T (PIG50), UBE2V2 (MMS2, UEV-2), UBE2S (E2EPF), UBE2C (Ubc10), His6-tagged UBE2W (Ubc 16), UBE2Q2, His6-tagged UBE2A, Non-tagged UBE2G2 (Ubc7), His6-SUMO tagged UBE2G2 (Ubc7), His6-tagged UBE2I (Ubc9), UBE2L6 (UbcH8), UBE2Z (Use1)

Recombinant Ubiquitin

Ubiquitin WT, Ubiquitin-K6, Ubiquitin-K11, Ubiquitin-K27, Ubiquitin-K29, Ubiquitin-K33, Ubiquitin-K48, Ubiquitin-K63, Ubiquitin-K0, Ubiquitin-K6R, Ubiquitin-K11R, Ubiquitin-K27R, Ubiquitin-K29R, Ubiquitin-K33R, Ubiquitin-K48R, Ubiquitin-K63R, Ubiquitin-K48R K63R


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  • 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)