PA770: PROTAC® In Vitro Ubiquitination Assay Kit




Proteolysis Targeting Chimeras (PROTACs) have emerged as novel therapeutics to target the undruggable proteome. PROTACs are heterobifunctional small molecules that both simultaneously bind to a target protein and a ubiquitin (Ub) E3 ligase. This subsequently leads to ubiquitination and degradation of the target protein. The In vitro ubiquitination kit has been developed to establish a high throughput approach that can accurately predict PROTAC efficiency by monitoring the protein’s intrinsic ability to get ubiquitinated. We offer this kit for three E3 ubiquitin ligases: Cereblon, VHL, and HDM2 to monitor PROTAC mediated ubiquitination for target of choice.

PROTAC In Vitro Ubiquitination Assay Kit Components

In Vitro ubiquitination Assay Plate 1 plate
10X Assay Buffer 1.2mL
E3 Mix (20X) 1 X 250µL
E1-E2-Ubiquitination Mix (20X) 1 X 250µL
Secondary Antibody (Anti-Mouse or Anti-Rabbit HRP conjugate) 60µL (100X)
Detection Reagent 1

Detection Reagent 2

DR1- 1mL

DR2- 1mL

Blocking Concentrate (5X) 5mL
Target Protein positive control BRD3 (20X) 20µL
Anti-BRD3 antibody (100X) 5µL
Anti-mouse HRP Conjugate (100X) 5µL

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Learn more about PROTACs here.

Additional information

PROTAC® In Vitro Ubiquitination Assay Kit

PA:770 Cereblon Kit, PA:770 VHL Kit, PA:770 MDM2 Kit

Secondary Antibody

Mouse, Rabbit


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  • Testing PROTAC and molecular glue (MGs) activity.
  • Compare between multiple PROTAC variants and establish predictive DC50 from UbMax.
  • Choose all three ligases for comparing PROTACs / MGs activity to demonstrate selectivity.
  • Test PROTAC substrate specificity and Isoform selectivity.