Immunoblot analysis of head microsomes fractionated by two phase affinity partitioning alone or by density gradient centrifugation followed by two phase affinity partitioning at 5.7% PEG/Dextran. A - Fractionation by two-phase affinity partitioning alone. The yield of Nervana is significantly improved (compare to Figure 4A.) However, BiP is still present in the ConA fraction indicating that 2PAP alone at the 5.7% concentration is still insufficient to produce high purity PM. Labeling: same as figure 2. Loading: All samples have equivalent loading, except P100, which was 1/5th of the others. B - Fractionation by two phase affinity partitioning following an initial density gradient fractionation. The Nervana is still found in the ConA sample, but this fraction no longer contains detectable BiP. Small amounts of residual ATP synthase are still present. Labeling: same as figure 4. Loading: Equivalent amounts of all fractions were loaded.
Khanna et al. BMC Genomics 2010 11:302 doi:10.1186/1471-2164-11-302