Beef heart cytochrome c oxidase was initially delipidated by incubation of the complex in 5% Triton X-100 followed by separation of the resulting detergent-protein complex from the detergent-lipid mixed micelles by sedimentation through a glycerol gradient containing 1% Triton X-100. After this treatment, the complex contained 2-3 mol of diphosphatidylglycerol (DPG) per heme au3. Further .delipidation could be achieved by a second 5% Triton X-100 incubation and a second glycerol gradient. After the second Triton X-100 treatment, the complex contained only 1-2 mol of DPG per heme a q .
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