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Reaction was performed in 25 mM HEPES, 100 mM NaCl, 10 mM MgCl 2 , 5 mM ATP, pH 7.5 with 2 µM UBE2D, 2 µM UBE2G1, 0.2 µM UBA1, and was initiated by adding 60 µM ubiquitin at room temperature

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