Organellar distribution of 1D9- and anti-PMP70-immunoreactive proteins in enriched peroxisomal rat liver fractions. Percoll-, Nycodenz- and Percoll/Nycodenz-purified peroxisomal fractions were processed for pre-embedding double immunoelectron microscopy using anti-Arf 1D9 (18 nm gold grains) and anti-PMP70 (12 nm gold grains) antibodies (see legend to Figure 3). Epon-embedded fractions were quantitatively analyzed for the density of labeling on organellar membranes. The results are expressed as relative specific labeling (R.S.L.) versus percentage of total cell organelles. R.S.L. is hereby defined as the percentage of total gold particles present on a particular organelle divided by the corresponding percentage of total organelles. R.S.L. values greater than one indicate enrichment of the labeling in that specific fraction. The combined total number of cell organelles (x-axis) and gold particles (y-axis) is indicated. Note that the purity of the peroxisomal Percoll, Nycodenz, and Percoll/Nycodenz fractions was 60%, 70%, and 77%, respectively. Abbreviations: PO, peroxisomes (vesicular structures labeled with anti-PMP70 antibodies); MT, mitochondria (double membrane-bound organelles with cristae); O, other vesicles (vesicular structures which could not be unambiguously identified as peroxisomes or mitochondria).
Anthonio et al. BMC Cell Biology 2009 10:58 doi:10.1186/1471-2121-10-58