Additional file 1.
Golgi and post-Golgi proteins in head microsomes fractionated by density gradient centrifugation followed by two phase affinity partitioning at 5.7% PEG/Dextran. A - Fractionated microsomes prepared from heads and probed for the Golgi protein Lava lamp. Two isoforms of Lava lamp are detected (arrows) at ~170 kDa and ~315 kDa. On average Golgi membrane is heavier than the peak ER fractions as expected ); double headed arrow; see Figure 3), but some overlap is seen especially with the larger isoform which has a bimodal distribution. B - Fractionated microsomes prepared from heads and probed with anti-Horseradish Peroxidase (HRP). The epitopes recognized by anti-HRP depend on the presence of N-glycan core α1,3-linked fucose  and thus detects proteins in trans-Golgi and post-Golgi compartments. The prominent epitope at 42 kDa is thought to be our PM marker Nervana . Trans- and post-Golgi proteins detected by anti-HRP extend from the Golgi fractions through to the lightest region of the gradient as seen with the fully glycosylated Nervana isoforms (see Figure 3A). C - Fractionation by two phase affinity partitioning following an initial density gradient fractionation and probed with anti-HRP. The most prominent band behaves the same way as Nervana (see Figure 6B) and probably is Nervana (see ). An overexposure of the final ConA eluate (ConA over) is included to show that other anti-HRP detectable proteins are also present in the PM fraction. Labeling: same as figure 3 for A and B and figure 4 for C. Loading: Equivalent amounts of all fractions were loaded.
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Khanna et al. BMC Genomics 2010 11:302 doi:10.1186/1471-2164-11-302