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After 48 h transfection of HEK293t cells with the HA-Sin1 plasmid, cells were lysed in CHAPS buffer (40 mM HEPES, pH 7.4, 120 mM NaCl, 1 mM EDTA, 0.3% CHAPS, 50 mM NaF, 5 mM sodium pyrophosphate (Na 4 P 2 O 7 )
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Usually, at physiological pH (7.0–7.4), the value of pKa is ~8.5
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