RP - In vitro ubiquitination assay (»Add to Infobox)

Di Fiore Pier Paolo, Polo Simona

Pier Paolo Di Fiore's Group, Simona Polo's group

Modification with ubiquitin (Ub) has been reconstituted in vitro for many known Ub targets. Assays usually involve incubation of bacterially expressed targets with recombinant or purified enzymes, Ub and ATP. Ub modification results in a target mobility shift of 8kDa, and is detected by anti-Ub immunoblot. The efficiency of in vitro ubiquitination assay depends on the quality of the enzymes involved. Here is described the assay developed in our laboratory.

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Di Fiore Pier Paolo (Principal Investigator / Pier Paolo Di Fiore's Group)

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