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. Author manuscript; available in PMC: 2017 Aug 3.
Published in final edited form as: Anal Chem. 2017 Mar 29;89(8):4428–4434. doi: 10.1021/acs.analchem.6b03675

Figure 1.

Figure 1

General workflow for isolation and detection of branched Ub chains by UbiChEM-MS. Cells are grown under different treatments and lysed under nondenaturing conditions. Clarified lysates are subsequently incubated with a resin bearing ubiquitin binding domains, and the isolated chains are then subjected to minimal trypsinolysis under nondenaturing conditions yielding Ub1–74 fragments containing Gly-Gly modifications. These products are enriched for using reverse-phase chromatography and characterized by high resolution mass spectrometry.