DUB Profiling Services

Boston Biochem offers customized in vitro profiling of your deubiquitinating enzyme (DUB), DUB substrate, and/or DUB activator or inhibitor. We can characterize your known DUB or confirm your putative DUB by testing it against all of the classic DUB substrates and inhibitors currently available. Our scientists will assay the activity of your DUB with any or all of our available substrates and inhibitors or test your inhibitor, substrate, or activator against the most comprehensive list of full-length DUBs available.

Ubiquitin specificity can be determined by assaying for cross hydrolysis or inhibition using our extensive list of Ubiquitin-like modifier (UBL) substrates and inhibitors.  If applicable, we can customize the appropriate components and information as a kit for you to test your enzyme or inhibitor in-house.

Summary

  • Selectivity and specificity of DUB substrates
  • Most comprehensive substrates available to monitor activity of all DUBs
  • Potency of compound vs. existing DUB panel
  • Single concentration or IC50 determination
  • Test activation or interaction of your protein with known DUBs
  • Customizable panel - over 40 DUBs to choose from
  • Most comprehensive substrates available to monitor activity of all DUBs

 

Contact us for further details

Phone: 617-576-2210

Email: contractservices@bostonbiochem.com

DUB Profiling request

Characterization of your DUB Enzyme

Substrate Specificity

Not all DUBs hydrolyze the same substrates in vitro. Some prefer small ubiquitin C-terminal leaving groups (Ubiqui-Fluors) while others will only hydrolyze the real isopeptide bonds in ubiquitin chains. A group exists that can do both at various efficiencies and those efficiencies can be evaluated and quantitated. Of the DUBs that hydrolyze chains, they can also have linkage specific activity. We can quickly and efficiently match your DUB to the appropriate substrate and if multiple substrates are viable, we can determine which is optimal for your research application or assay format. Other ubiquitin-like substrates for the study of NEDPs, SENPs, etc. can be tested for cross hydrolysis with your enzyme for complete characterization.

Inhibitor Specificity

Another important confirmation of a DUB enzyme is the ability to block activity with classic DUB inhibitors. Once an optimal in vitro substrate is chosen and assay format is established, we will test our known DUB inhibitors versus your enzyme for further characterization. Other ubiquitin-like inhibitors for the study of NEDPs, SENPs, etc. can be tested for cross inhibition with your enzyme for completion of characterization.

Characterization of your Inhibitor, Activator or Substrate

Inhibitor

Boston Biochem has expertise that allows you to test your protein, peptide or small molecule inhibitor with any or all of our available DUBs. Critical to this analysis is the use of appropriate substrates and assay formats for each DUB to generate interpretable data. We have assays up and running for every DUB in our portfolio. If you are studying a DUB that is not in our range, we will work with you to develop the appropriate assay to properly assess inhibitory activity. All inhibition assay formats will be confirmed with positive control DUB inhibitors.

Activator

Test your small molecule, peptide, or protein for the ability to increase hydrolysis rates of any or all of our DUBs versus the appropriate substrates.

Substrate

Boston Biochem will determine what DUBs hydrolyze your substrate via appropriate assay. If there are multiple DUBs that show activity against your substrate we will determine efficiencies and rates for the positively identified DUBs.

Similar services available for other UBL Deconjugating Enzymes