A kind of extraction method of enoki mushroom mitochondria
An extraction method and mitochondrial technology, applied in the field of microorganisms, can solve the problems of mitochondrial inner membrane damage, mitochondrial outer membrane pollution, poor mitochondrial purity, etc., and achieve the effects of short time consumption, good integrity, functional activity, and high functional activity.
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Embodiment 1
[0054] Immerse the sample in mercaptoethanol solvent, soak at 32°C for 20min, centrifuge at 500×g for 3min, discard the supernatant, resuspend the pellet in sorbitol, add 35mg·g helicase -1 (according to the weight of the precipitate obtained in the above steps), incubate at 37°C for 1h; centrifuge at 500×g for 3min, discard the supernatant, wash the precipitate three times with PBS, and collect the precipitate by centrifuging at 500×g for 3min at 4°C; add 1.5 Resuspend the pellet in ml ice-cold Mito-Cyto Buffer (derived from the mitochondrial-cytoplasmic protein preparation kit purchased from Beijing Pulilai Gene Technology Co., Ltd.), and the volume ratio of the pellet mass to Mito-Cyto Buffer is 1g: 5ml, 4°C, centrifuge at 800×g for 5 minutes, collect the supernatant and centrifuge again at 800×g for 5 minutes, collect the supernatant and centrifuge again at 10,000×g for 10 minutes, the sediment at the bottom of the tube is thick mitochondria. The pellet was resuspended wit...
Embodiment 2
[0056] The steps are the same as in Example 1, except that the enzymatic hydrolysis time is 55 minutes.
Embodiment 3
[0058] The steps are the same as in Example 1, except that the enzymatic hydrolysis time is 65 minutes.
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