Mitochondrial Isolation Kit

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Size	Unit Price	Belgium	Japan*	Quantity
1KIT	336,00 €	1	19

*Stock available in Belgium will be delivered in 1 to 3 days
*Stock available in Japan will be delivered in 1 to 2 weeks (excludes regulated items and dry ice shipments).

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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

• 1. Collect cells via centrifugation and carefully remove supernatant. Use a cell scraper for detachment of adherent cells.
  2. Resuspend cells in Mitochondrial Isolation Reagent A and vortex for 5 seconds. Add 400 μL for every 1 x 10⁷ cells.
  3. Incubate on ice for 2 minutes.
  4. 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.
  5. Incubate on ice for 5 minutes, vortexing once per minute.
  6. 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.
  7. Centrifuge at 700 x g for 10 minutes at 4°C.
  8. Transfer the supernatant to a new tube and centrifuge at 12,000 x g for 15 minutes at 4°C.
  9. Transfer the supernatant (cytosolic fraction) to a new tube and add 300 μL of Mitochondrial Isolation Reagent C to the pellet (mitochondria).
  10. Centrifuge at 12,000 x g for 5 minutes at 4°C and remove supernatant.
  11. Mitochondria (pellet) are now ready to be used in further experiments.

Storage

• Store at 4 °C.