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. 2010 Feb 26;9(7):1514–1532. doi: 10.1074/mcp.M900421-MCP200

Fig. 3.

Fig. 3.

Immunoblot and MS/MS-SILAC analyses of chloroplast and mitochondrial samples show low contamination levels. A, immunoblot analysis of 35 μg of total protein from chloroplast and mitochondrial samples from AR and AN conditions. Detection using antibodies against chloroplast (LhcbM6 and PsaD) and mitochondrial (CoxIIB) marker proteins shows low, but constant contamination of mitochondrial proteins in the chloroplast samples. On the other hand, the mitochondrial samples contain significantly less chloroplast protein contamination. B, MS/MS-SILAC analysis of mitochondrial samples mixed with labeled chloroplasts reveals detection of mitochondrial proteins in the heavy isotope range. For the ATP2, the ratio of mitochondria to chloroplasts is 10.895 (S.D. 3.545). A spectrum for a peptide belonging to this protein is shown.