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E3 ligases assays

Creative Biogene is offering assays covering the most comprehensive E3 ligases. With years of experience, we have successfully developed the platform for commercial and research application allowing researchers to screen against E3 ligases in either HTS or non-HTS formats.

Creative Biogene can provide a wide range of assays for E3 ligases, including E3 ligase expression and purification, E3 ligase in vitro activity assays, binding affinities of E3 ligase inhibitors, and E3 in cells substrate ubiquitylation assays.

The E3 ligase family is the largest family in ubiquitin signaling pathway with about 700 members identified or predicted to possess ligase activities. E3 recruits an E2 ubiquitin-conjugating enzyme that has been loaded with ubiquitin, then recognizes a protein substrate, and assists or directly catalyzes the transfer of ubiquitin from the E2 to the protein substrate (Degui, et al. 2016)

E3 ubiquitin ligases can be classified into three subfamilies. First is RING E3s, which act as scaffolding molecules to bring ubiquitin-charged E2 enzymes in close contact with their substrates. Second is HECT E3s, which catalyze the transfer of ubiquitin to their own cysteine residues and subsequently to substrates. And the third subfamily, RING-Between-RING (RBR) E3s, which include PARKIN and ARIH1, and mechanistically behave as hybrids between RING and HECT.

Since E3s are a large family of enzymes that has distinct catalytic mechanisms, targeting E3s is anticipated to yield better specificity, less toxicity and be a more superior option (Xiaodong, et al. 2016). Therefore they are very promising potential therapeutic targets for drug discovery.

Fig.1 Summary of the ubiquitin system (Xiaodong, et al. 2016)Fig.1 Summary of the ubiquitin system (Xiaodong, et al. 2016)

The basic services that Creative Biogene offers for E3 ligases are as follows:

  1. For E3 ligase expression and purification assays, we are providing a broad list of E3 ligases.
  2. For E3 ligase in vitro activity assays, we offer ELISA-based autoubiquitylation or substrate ubiquitylation assay and TR-FRET autoubiquitylation or substrate ubiquitylation assay.
  3. For binding affinities of E3 ligase inhibitors, we offer both thermal shift assay and surface plasmon resonance assay (SPR).
  4. For E3 in cells substrate ubiquitylation assays, we offer ELISA-based substrate ubiquitylation assay and AlphaLISA-based substrate ubiquitylation.

Creative Biogene offers assays from ligase expression to low- or high-throughput in vitro assays to cellular assays, which allows you to confirm compound activity against your target ligase in multiple contexts and formats. Our talented scientists with abundant expertise will work close with you to accelerate your ligase project with high quality and efficiency. Creative Biogene has confidence in speeding your ligase target drug discovery project with fast turnaround time and the most competitive price.

If you have any special requirements in E3 ligases assays services, please feel free to contact us at or 1-631-626-9181. We are looking forward to working together with your attractive projects.


  1. Xiaodong, H. and Vishva, M. D. (2016) ‘Drugging the undruggables: exploring the ubiquitin system for drug development’, Cell Research, 26, 484-498
  2. Degui, W., Yingxia, T., Dong, W., Yuhong, J., Haitao, N., Kun, X., Yanfeng, S. (2016) ‘DNA Damage-Induced Foci of E2 Ubiquitin-Conjugating Enzyme are Detectable upon Co-transfection with an Interacting E3 Ubiquitin Ligase’, Biochem Genet, 54(2):147-57

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