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Western blot analysis of an in vitro Rsp5 ubiquitination reaction with CTD (+CTD lanes) or lacking CTD in the presence of Rup1-TAP (+Rup1 lanes) purified from a strain lacking UBP2. Reactions were incubated from 5 minutes to 4 hours as indicated and stopped by the addition of sample loading buffer. Ubiquitinated and unmodified forms of GST-tagged Rsp5 were detected with anti-GST antibody.
