Samples were prepared in 1x sample loading buffer [50 mM Tris-HCl pH 6.8, 2% SDS, 10% glycerol, 1% β-mercaptoethanol, 12.5 mM EDTA and 0.02% bromophenol blue] and proteins were separated by SDS/PAGE (10% or 12% Bis-Tris gel; Life technology), and then transferred to a PVDF membrane for blotting (BioRad Laboratories).
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