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The 0.4 ml of supernatant was added to 0.5 ml of 1.25 M NaH 2 PO 4 and 0.1 ml of 100 mM NEM dissolved in 125 mM NaK phosphate buffer (pH 7.5)

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Protein (75 g) was loaded on an 8% T, 2.5%C polyacrylamide gel