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Ubiquitination of GFP-tagged proteins

GFP-Trap® for the analysis of candidate ubiquitinated proteins under harsh conditions.

In our approach to analyze ubiquitination of individual candidate proteins in living cells we co-transfect the GFP-tagged protein with FLAG-tagged ubiquitin. Due to strong binding of the GFP-Trap in combination with covalent ubiquitination, highly specific washing steps under denaturing conditions (8 M Urea and 1% SDS) are possible to remove unspecifically bound proteins.