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For the purposes of immunoblotting, isolated beads were reconstituted in 50 L of lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, protease inhibitors), incubated on ice for 10 min and lysates cleared by centrifugation at 17,000 g for 10 min

In addition, the applicant stated that the Johnson et al