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. 2013 Sep 19;8(9):e75372. doi: 10.1371/journal.pone.0075372

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© 2013 Lam, Emili

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Western blot showing Ubp2 reversal of mobility shifted Rsp5-Ub species. Ubiquitination reactions were performed in the presence of substrate (CTD) to stimulate Rsp5 auto-ubiquitination (+ub rxn). Ubp2-TAP (+Ubp2), buffer only (-Ubp2), and/or the DUB inhibitor ubiquitin aldehyde (+ub-ald) were then added, and the reactions allowed to incubate for a further 2 hours (+incub). Ubiquitinated and unmodified forms of GST-tagged Rsp5 were detected with anti-GST antibody.