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P2 was suspended in 10 vol of 0.32 M sucrose and 10 ml of the solution layered on a 0.8 and 1.2 M two-step discontinuous gradient, which was centrifuged at 61,000 x g for 2 h to separate the synaptosomes from the mitochondria (which forms the pellet)

R., Busby-Whitehead, J., Tsiao, C., Stewart, K