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