50 mM HEPES (pH 7.5), 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 10% glycerol, and 10 mM sodium pyrophosphate. Following reagents were added just before lysis at the indicated concentrations: 2 mM sodium orthovanadate, 10 mM sodium fluoride, 1 mM PMSF, 5 μg/ml leupeptin, 5 μg/ml pepstatin and 5 μg/ml aprotinin.
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