After adding 2X sample buffer (100 mM Tris pH 6.8, 200 mM DTT, 4% SDS, 0.2% Bromophenol blue, 20% glycerol), the PSD samples were then boiled at 95°C for 5 min and 30 µg of each sample was loaded for separation on an 4–12% SDS-PAGE gradient gel (Invitrogen).
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