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Then, the strips were equilibrated in equilibration buffer containing 2% (w/v) SDS, 1% (w/v) DTT, 50 mmol/L Tris-HCl, pH 8.8, 20% (v/v) glycerol, 6 mol/L urea, 0.002% (w/v) bromophenol blue and 1.5% (w/v) iodoacetamide for 15 min at room temperature
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