Abstract: Objective To compare different methods for extracting mitochondria and its effects on mitochondria content and their activity. Methods Pulilai kit, sucrose gradient centrifugation and Pierce mitochondria extraction kit were used to extract mitochondria and quantify their concentration, activity and contamination. Results Pulilai kit yielded higher concentration of mitochondria proteins than two other methods(P<0.05) when the same amount of cells were used. Western blotting was used to evaluate the amount of cytosolic protein contaminant and mitochondria protein cytochrome c from mitochondria fraction. The result suggests that using Pulilai kit can yield lower concentration cytosolic protein contaminant and higher concentration of cytochrome c from mitochondria fraction than the other two methods. At the same time, ATP content of mitochondria extracted by the method of Pulilai kit was the greatest. Conclusion Pulilai kit is better than sucrose gradient centrifugation and Pierce mitochondria extraction kit for mitochondria extraction.

Key words: mitochondria, mitochondrial extraction, mitochondrial activity