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.
