Research reportA rapid Percoll gradient procedure for isolation of synaptosomes directly from an S1 fraction: homogeneity and morphology of subcellular fractions
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2021, NeuropharmacologyCitation Excerpt :Frozen striata from 30-day-old male naïve wild-type, mGlu2−/− and mGlu3−/− mice were used for the assessment of DA release in synaptosomes. Synaptosomes were isolated as described previously (Dunkley et al., 1988; Franklin and Taglialatela, 2016). Briefly, the tissue was homogenized in 10 vol of 0.32 M sucrose, buffered to pH 7.4 with tris-(hydroxymethyl)-amino methane (TRIS, final concentration 0.01 M) using a glass/Teflon tissue grinder (clearance 0.25 mm); the homogenate was centrifuged at 1000×g for 5 min to remove nuclei and debris, and the supernatant was gently stratified on a discontinuous Percoll gradient (6%, 10% and 20% v/v in Tris-buffered sucrose) and centrifuged at 33,500 g for 5 min.
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