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After centrifugation of lysates (14,000 rpm, 10 min, 4 C), the 30 L of supernatant was transferred to new 1.5-mL tube with 10 L of 4SDS sample buffer (200 mM Tris-HCl pH 6.8, 8% SDS, 40% glycerol, 100 mM DTT, 0.04% bromophenol blue
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