How should Accutase be used for cell detachment?
Accutase can be used directly, without dilution, following this short protocol:
- Wash the cells with sterile PBS (without Ca²⁺/Mg²⁺) or Hepes‑BSS
- Add Accutase to the culture vessel (example: 2-3 ml per 25 cm² or 8 ml per 75 cm² TC flask)
- Incubate for 5-20 min at room temperature, or 37°C for faster detachment
- Once the majority of cells has detached, collect the cell suspension, dilute it with fresh medium
- Centrifuge and resuspend the cell pellet in fresh culture medium
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