Isolation of Mitochondria and SDH Assay Assignment
Winter 2021
Isolation of Mitochondria
1. Why was it important to use the ends of the florets of the cauliflower rather than the stalks?
(2 marks)
2. What is differential centrifugation? Explain how it can be used to separate cellular organelles.
(3 marks)
3. Explain why it is essential during the isolation process to keep the homogenate and isolates cold.
(3 marks)
4. Both the isolation buffer and assay buffer that was used contained 0.3M mannitol. What was the purpose of including mannitol at this concentration? (3 marks)
5. was the purpose of the catalase test? (2 marks)
Succinate Dehydrogenase Assay
6. What is succinate dehydrogenase (SDH)? How is it useful in determining mitochondria activity in your collected solutions? (3 marks)
7. How is 3,4-dichlorophenol-indophenol (DCPIP) used to measure SDH activity? (3 marks)
8. Why is it essential to add sodiumazide to your assay solution? What is the purpose of including the sodium succinate solution? (4 marks)
