and 10 mM MgCl2at 37 °C overnight. To purify ubiquitinated Eos3.2 from free ubiquitin chains, the reaction mixture was incubated with Ni-NTA resin, eluted with 300 mM imidazole, and run over a Superdex 200 size-exclusion column equilibrated in the SEC Buffer. Fractions with long ubiq...
Approximately 100 μg of cell lysates were put aside for complete proteome analysis while the remaining material was mixed with 5 ml of Ni2+-NTA agarose beads (Qiagen) pre-equilibrated in binding buffer, and the slurry was incubated at 4 °C overnight. Beads were collected and washed...
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