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The daily trends for a 55 years old man and woman, waking up at 7 am, are shown in Fig

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The wells were blocked with DPBS (pH 7.4)/4% (w/v) skimmed milk powder (Sigma-Aldrich) (PBS/M) (250 L/well) for 1 h at room temperature and washed three times with PBS/0.05% (v/v) Tween 20 (PBS/T) at either pH 5.5 or pH 7.4