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After incubation for a further 2 h at 4 C, the beads were washed three times in NP-40 buffer (20 mM Tris-HCl pH 8.0, 137 mM NaCl, 1% NP-40, 10% glycerol, 2 mM EDTA and 1% protease inhibitor cocktail) and centrifuged at 100 g at 4 C for 1 min

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