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. Author manuscript; available in PMC: 2011 Jul 23.
Published in final edited form as: Biochem Biophys Res Commun. 2010 Jun 25;398(2):272–277. doi: 10.1016/j.bbrc.2010.06.077

Figure 1. Intact mitochondria and inner membrane preparations are sufficiently pure for sphingolipid determination.

Figure 1

Microsomal and lysosomal contamination of the IFM was monitored by measuring protein disulfide isomerase (PDI) and N-acetyl-β-D-glucosaminidase (NAG) activity, respectively. (A) Western blot analysis shows no detectable PDI in IFM preparations. n=3; (B) The percentage of NAG in IFM and SSM were ≤1.5% of that found in cardiac tissue homogenates. N=3; (C) Optimal digitonin levels to solubilize mitochondrial outer membranes from the IMM were determined. Western blot analysis showed successively greater removal of VDAC, a protein marker of mitochondrial outer membranes, with increasing digitonin levels added; however, COX IV (an inner membrane marker) remained constant regardless of digitonin levels. The blot shown is typical of four preparations.