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The immunoprecipitated mixture was then washed sequentially with a Triton dilution buffer (1% Triton X-100, 2 mM EDTA pH 8, 150 mM NaCl and 20 mM Tris-HCl pH 8.1), mixed micelle wash buffer (1% Triton X-100, 5 mM EDTA pH 8, 150 mM NaCl, 20 mM Tris-HCl pH 8.1, 5% sucrose, 0.2% NaN 3 and 0.2% SDS), buffer 500 (0.1% w/v deoxycholic acid, 1 mM EDTA pH 8, 50 mM HEPES pH 7.5, 500 mM NaCl, 1% Triton X-100, 0.2% NaN 3 ) and a LiCl wash buffer (0.5% w/v deoxycholic acid, 1 mM EDTA pH 8, 250 mM LiCl, 0.5% v/v NP-40, 10 mM Tris-HCl and 0.2% NaN 3 )

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