几种不同提取线粒体的方法对线粒体含量及活性的影响

Comparison between Different Methods to Extract Mitochondria and Its Effects on Mitochondria Content and Activity

目的比较几种不同的提取线粒体的方法对线粒体蛋白浓度及活性的影响。方法采用蔗糖密度梯度离心法、普利莱试剂盒提取方法和Pierce线粒体提取试剂盒方法分别提取线粒体,测定提取得到的线粒体蛋白浓度、污染情况和活性。结果同样数量的细胞采用普利莱法提取得到的线粒体蛋白浓度高于其他2种方法所提取的线粒体蛋白浓度(P<0.05);采用Westernblotting的方法测定提取的线粒体发现采用普利莱法提取得到的线粒体中胞质蛋白污染较其他2种方法提取得到的线粒体少,线粒体纯度高;测定线粒体中细胞色素C和ATP含量后发现同样采用普利莱法提取得到的线粒体活性最高。结论普利莱法提取线粒体的方法优于蔗糖密度梯度离心法和Pierce线粒体提取试剂盒方法。

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 c...