Succinate Dehydrogenase Lab Report

The purpose of this experiment was to determine the concentration of succinate dehydrogenase in six fractions made by students. Succinate dehydrogenase is an enzyme located in the mitochondrial membrane and aids in cell respirations and enzyme generation. In order to conduct this experiment, students cut a liver and through a process created six fractions. Each of the six fractions was moved to a cuvette with solutions containing different volumes and recoding the absorbance every five seconds for four minutes. Once the absorbance was obtained for each, we determined that cuvette E (0.0012) had the least activity per milliliter while cuvette B had the most activity per milliliter 0.0138 (Table 2). The higher the activity per milliliter the higher the concentration of succinate dehydrogenase—which also means more oxidation. As it turns out, the more the fractions spun in the centrifuge, the lower the concentration of succinate …show more content…
The T.A then spun the centrifuge at 1000rpm for 10 minutes—in order for the cell remains and nuclei to form pellet. After, the supernatant was placed in a centrifuge tube. Students added 4.0ml of homogenization buffer to the pellet and re-suspended it using the Vortex. 1.5ml of this solution was placed in two different microcentrifuge tubes labeled “nuclear.”
Next, we placed the tube of supernatant in the ultracentrifuge and spun it at 10000rpm for 20 minutes. Students then added 1.5ml of the supernatant in two different microcentrifuge tubes labeled “post- mitochondria.” The rest of the supernatant was placed in a test tube labeled “post mitochondria” for use in activity A4. The pellet was re-suspended after 4.0ml of homogenization buffer was added. 1.5ml of this solution was placed in two different microcentrifuge tubes labeled “mitochondria.” The remaining solution was appropriately thrown