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Target Ubiquitination

Being a certified contract research organization (CRO) with GLP facility, Creative Biolabs is experienced in offering one-stop PROTAC service. To support institutes and pharmaceutical industry for therapeutic PROTAC small molecule drugs, we offer various services to assess the target protein ubiquitination targeted by PROTAC candidates.


As a rapidly emerging alternative therapeutic strategy, PROTAC has the potential to address many of the current challenges faced in modern drug development approaches. PROTAC employs small molecules composed of a target protein ligand, a linker, and an E3 ligase ligand. Following “target protein-PROTAC-E3 ligase” ternary complex formation, the exposed lysines on the target protein will be ubiquitinated by E3 ligase complex. Upon poly-ubiquitination of the target protein, it will be recognized by proteasome and degraded into amino acids and small peptides (see Fig.1).

Schematic of the PROteolysis TArgeting Chimera (PROTAC) technology. Fig.1 Schematic of the PROteolysis TArgeting Chimera (PROTAC) technology. (Neklesa, 2017)

Introduction of Target Ubiquitination Service

As per the above PROTAC principles, we can draw that E3 ligase complex-mediated ubiquitination for target protein is crucial determinant for target protein degradation. To evaluate the target protein ubiquitination efficacy caused by PRORAC candidates, Creative Biolabs has developed a series of different approaches to facilitate your PROTAC research. To avoid false positive, we add the controls for each assay, including proteasome or ubiquitination inhibitors. Our approaches are listed below, including but not limited:

  • Mass Spectrometry (MS)
  • MS-based proteomics has become a powerful technique for the analysis of protein ubiquitination. It measures the mass-to-charge ratio (m/z) of ions to identify and quantify ubiquitination in simple and complex mixtures. Creative Biolabs offers MS-based approach to characterize the target protein ubiquitination after treated with PROTACs. We ensure high efficiency and high sensitivity.

  • Western Blot (WB)
  • WB is a widely used technique in molecular biology to detect specific proteins via corresponding antibodies. Creative Biolabs offers a series of processes - cell lysis, immunoprecipitation, washing, and anti-ubiquitin immunoblotting to evaluate the target protein ubiquitination treated with PROTAC. The degree of ubiquitination can be estimated by comparing the ratio of ubiquitinated/unmodified target protein in different experimental conditions.

(A-D) Western blot ubiquitination results (Ottis, 2017); (E) Workflow for enriching ubiquitylated proteins Fig.2 (A-D) Western blot ubiquitination results (Ottis, 2017); (E) Workflow for enriching ubiquitylated proteins (Heap, 2017).

Highlight Features of Our Services

  • High efficiency
  • Assay design plus conduction
  • Accurate results
  • Superior after-sale service

As the important signal prior to target protein degradation, target protein ubiquitination is a crucial index to indicate the degradation efficacy. Creative Biolabs is here to serve both MS and WB approaches to evaluate the ubiquitination efficacy, and we are committed to choosing the best optimal assay to maximize the success of your projects. Please contact us for any questions or further communication.


  1. Ottis, P.; et al. Assessing Different E3 Ligases for Small Molecule Induced Protein Ubiquitination and Degradation. ACS Chem Biol. 2017, 12(10), 2570-2578.
  2. Heap, R.E.; et al. Mass spectrometry techniques for studying the ubiquitin system. Biochem Soc Trans. 2017, 45(5), 1137-1148.
  3. Neklesa, T.K.; et al. Targeted protein degradation by PROTACs. Pharmacol Ther. 2017, 174, 138-144.
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