Brains from male Sprague Dawley rats were suspended in M-PER, Mammalian Protein Extraction Reagent, with protease inhibitor cocktail and homogenized using a Dounce tissue grinder. The tissue lysate was adjusted to 2mg/mL in SDS PAGE sample buffer (40 mM Tris-HCl pH 6.8, 1% SDS, 50 mM DTT, 7.5% glycerol, 0.003% bromophenol blue) and heated for 10 min at 70°C.
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