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That explanation reflects the timing of the process

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The membranes were washed in TBS-T (50 m M Tris–HCl, pH 7.6, 150 m M NaCl, 0.1% Tween 20) for 30 min and incubated with an appropriate Horseradish peroxidase-conjugated secondary antibody (1:5000 dilution) for 2 h at room temperature