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. 2012 May 24;7(5):e37378. doi: 10.1371/journal.pone.0037378

Figure 1. ABCB6 is not present in the purified mitochondrial fraction of K562 cells.

Figure 1

Cellular membranes were separated by differential centrifugation (A); mitochondria were isolated using anti-TOM22 magnetic beads (B) as described in Materials and Methods. Following separation by SDS-PAGE, organelle membranes were transferred to a PVDF membrane, which was incubated in antibodies recognizing specific organelle markers and ABCB6. A. Cellular fractions corresponding to total cell lysate (lane 1); nuclei and intact cells (lane 2); mitochondrial pellet of the 8000×g centrifugation (lane 3); pellet of 12.000×g centrifugation (lane 4); pellet of 20.000×g centrifugation (lane 5). B. Isolation of mitochondria using TOM22 immunobeads. Total cell lysate (lane 1); fraction bound to the beads after repeated washing steps (lane 2); pellet of the flow through (lane 3).