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Then, 20 L of pFeSAN (1 mg/mL) and 20 L of TMB (50 mM DMSO) were sequentially added into HAc-NaAc buffer (0.1 M pH 4) and the final volume was fixed to 1 mL, which contained different amounts of Hepa 16 or AML-12 (1000, 10,000, 50,000, 100,000, 200,000, 300,000, 500,000 or 800,000)