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On the next day, after the membrane was washed with TBST for five times, HRP-conjugated secondary antibody (Thermo Scientific) was applied for 1.5 h at room temperature
Months earlier, it abandoned another oral obesity drug that showed concerning liver effects in a trial
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A80-229AP, Bethyl Laboratories, Inc.), diluted in PBS/T/M (pH 5.5 or pH 7.4), was added for 1 h at room temperature, followed by a washing step and detection with 1 mg/mL (100 L/well) of p -Nitrophenyl Phosphate Substrate (Sigma-Aldrich) dissolved in diethanolamine buffer (pH 9.8) (Sigma-Aldrich)
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