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Excess DTT and imidazole were removed on a PD-10 desalting column (Merck), and the reduced proteins were eluted with 3.5 mL ice-cold assay buffer containing 100 mM Tris/HCl, 1 mM EDTA, pH 8.0

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Taken together, FM subjects have decreased isovaleric acid concentrations compared to HC, but these changes did not correlate with FM symptoms