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After incubation, active Ras was precipitated using glutathione-agarose beads, washed, and eluted in SDS sample buffer

After 1 min, the mitochondria-associated radioactivity was transferred to 0.4-mL centrifuge tubes (USA Scientific, Ocala, FL) containing a 100-L layer of oil consisting of 1:37.5 n-dodecane:bromododecane (Sigma) and placed into a Beckman E centrifuge