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Mitochondrial Isolation Kit
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Product Information
- Components : Mitochondrial Isolation Reagent A, Mitochondrial Isolation Reagent B, Mitochondrial Isolation Reagent C
- Size : 30 tests
Description
- This product can be used to quickly and easily isolate mitochondria from cultured mammalian cells and tissue. Isolated mitochondria can be used in downstream analyses.
Directions for Use
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- Collect cells via centrifugation and carefully remove supernatant. Use a cell scraper for detachment of adherent cells.
- Resuspend cells in Mitochondrial Isolation Reagent A and vortex for 5 seconds. Add 400 μL for every 1 x 107 cells.
- Incubate on ice for 2 minutes.
- Add Mitochondrial Isolation Reagent B to cell suspension and vortex for 5 seconds. Use 5 μL for every 400 μL of suspension from step 2.
- Incubate on ice for 5 minutes, vortexing once per minute.
- Add Mitochondrial Isolation Reagent C and invert several times (do not vortex). Use a volume equivalent to that used of Reagent A in step 2.
- Centrifuge at 700 x g for 10 minutes at 4°C.
- Transfer the supernatant to a new tube and centrifuge at 12,000 x g for 15 minutes at 4°C.
- Transfer the supernatant (cytosolic fraction) to a new tube and add 300 μL of Mitochondrial Isolation Reagent C to the pellet (mitochondria).
- Centrifuge at 12,000 x g for 5 minutes at 4°C and remove supernatant.
- Mitochondria (pellet) are now ready to be used in further experiments.
Storage
- Store at 4 °C.
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