In-vitro ubiquitination of a purified recombinant protein is not working. Do you have any suggestions for me to try?

I have UBE1, UBE2K, E3 Ligase, E3 Ligase buffer, Ubiquitin and mgATP in the rxn. The concentrations are adapted from an in-vitro Ubiquitination kit.

07 March 2017 3,047 2 View