Determination of PP2Cm as a soluble protein located in mitochondrial matrix. (A) Immunoblot for PP2Cm,TOM40, and Timm13 following carbonate extraction using 0.1 M Na2CO3 (pH 11.0). S, supernatant; P, pellet from centrifugation after Na2CO3 treatment. (B) Immunoblotting for PP2Cm, Hsp60 (matrix protein marker) and Timm13 (IMS marker) following mitoplasting assay using sucrose concentrations as indicated to produce hypotonic conditions.T, total; P, pellet; and S, supernatant following centrifugation. Reproduced from (Lu et al., 2007) with permission from CSHL Press.