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The collected protein pellet was washed once with 1 ml of chilled (20 C) methanol via probe-tip sonication (5 s on, 20 s off, 20% A, four to six pulses) on ice, transferred to a 2-ml microcentrifuge tube and centrifuged at 4 C for 5 min at 6,500 g

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However, alkaline phosphatase can remove the phosphate from 4-phosphopantetheine to generate pantetheine