DUB Assay Technology — Principles and Application Guide
What are Deubiquitinases (DUBs)?
Deubiquitinases are proteases that hydrolyze isopeptide bonds between ubiquitin and target proteins (or within ubiquitin chains). The human genome encodes approximately 100 DUBs across seven families, with the ubiquitin-specific protease (USP) family being the largest.
Fluorogenic Substrate Assay Principle
The fluorogenic substrate method is the most commonly used DUB activity detection approach:
- Conjugate a fluorophore (e.g., AMC, Rho110) to the ubiquitin C-terminus
- DUB cleavage releases free fluorophore, dramatically increasing fluorescence
- Monitor fluorescence signal change in real-time using a fluorescence plate reader
Product Selection Guide
Ub-AMC: Classic substrate, cost-effective, suitable for routine DUB activity assays.
Ub-Rho110: High sensitivity, ideal for low-concentration enzyme detection and high-throughput drug screening.