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The mixture was incubated at room temperature for 15 minutes, then added with 2.5 L of Pantetheine 7-amino-4-trifluoromethylcoumarin substrate, such that the final concentration of recombinant human Vanin-1 was 62.5 M and the final concentration of Pantetheine 7-amino-4-trifluoromethylcoumarin substrate was 45 M in the 10 L reaction system
