Table 1

Relative yields of membrane compartments determined from marker enzymes.

Method

Enzyme

Microsomes

PM Pool

ConA


Affinity

ALP

100

0.55 ± 0.44


SDH

100

0.37 ± 0.15


CCR

100

0.37 ± 0.26


Combination

ALP

100

0.7 ± 0.14

0.4 ± 0.17


SDH

100

0.8 ± 0.07

0.25 ± 0.04


CCR

100

1.1 ± 0.52

0.1 ± 0.02


Gradient+2PAP (6.3%)

ALP

100

54.0 ± 23.4


SDH

100

30.0 ± 5.31


CCR

100

12.9 ± 1.7


Embryos were used as the starting material. Yields of Alkaline phosphatase (ALP; EC 3.1.3.1; PM marker), Succinate dehydrogenase (SDH; EC 1.3.5.1; mitochondrial marker) and Cytochrome c reductase (NADPH) (CCR; EC 1.6.2.4; ER marker). Because significant amounts of ConA elute in the final fraction, specific activities cannot be presented and these figures are raw activity yields, normalized to the activity present in the microsome pellets. Results represent mean ± 1 standard deviation of values obtained from three experiments after adjustment for differences in protein concentration in the microsomal input fraction. The addition of the pre-fractionation by density gradient centrifugation preferentially reduces the amount of contaminating ER in the ConA fraction.

Khanna et al. BMC Genomics 2010 11:302   doi:10.1186/1471-2164-11-302

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