Abstract This experiment studies the rate of activity of succinate dehydrogenase protein derived from the mitochondria of cauliflower cells. Succinate dehydrogenase, once active, is known to reduce the coenzyme flavin adenine dinucleotide (FAD) to FADH2. Succinate dehydrogenase and its coenzyme creates an enzyme complex (E-FADH2) which transfers electrons to coenzyme Q during the Krebs cycle. In this experiment, sodium azide will be added to the mitochondrial solution to prevent the transfer of electrons to natural electron acceptors, such as coenzyme Q. This will transfer the electrons to an artificial electron acceptor called 2,6-dinitrophenolindophenol. Hence, the activity of succinate dehydrogenase is measured by the change in
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Afterwards, the cauliflower, along with five grams of cold purified sand and forty milligrams of cold mannitol, were added to a cold mortar and ground with a pestle. The ground mixture is then filtered through four layers of cheese cloth. The light-brown liquid filtrate is accumulated in a beaker before it is placed into a 50ml centrifuge tube. After the tubes are balanced by weight, they are placed into the centrifuge for ten minutes at 600g and a temperature range from zero to four degrees Celsius. The solution is poured into another centrifuge tube to prevent any solids of the sand, silt, or remnants of small pieces of cauliflower to remain in the liquid containing the cauliflower cells. The postnuclear supernatant fluid is centrifuged at 2200rpm for 30 minutes and a temperature range from zero to four degrees Celsius to acquire the mitochondrial pellet. The postmitochondrial supernatant fluid is discarded, and seven milliliters of mannitol assay medium at room temperature is added to the pellet. The pellet is scraped and mixed with the mannitol to disperse the mitochondrial fraction evenly. Afterwards, the mitochondrial suspension was immediately put on ice to prevent any enzyme
Afterwards, the cauliflower, along with five grams of cold purified sand and forty milligrams of cold mannitol, were added to a cold mortar and ground with a pestle. The ground mixture is then filtered through four layers of cheese cloth. The light-brown liquid filtrate is accumulated in a beaker before it is placed into a 50ml centrifuge tube. After the tubes are balanced by weight, they are placed into the centrifuge for ten minutes at 600g and a temperature range from zero to four degrees Celsius. The solution is poured into another centrifuge tube to prevent any solids of the sand, silt, or remnants of small pieces of cauliflower to remain in the liquid containing the cauliflower cells. The postnuclear supernatant fluid is centrifuged at 2200rpm for 30 minutes and a temperature range from zero to four degrees Celsius to acquire the mitochondrial pellet. The postmitochondrial supernatant fluid is discarded, and seven milliliters of mannitol assay medium at room temperature is added to the pellet. The pellet is scraped and mixed with the mannitol to disperse the mitochondrial fraction evenly. Afterwards, the mitochondrial suspension was immediately put on ice to prevent any enzyme