PBS-EDTA (0.5 mM, enzyme-free dissociation) recipe
Detaches cells without trypsin when surface proteins must be preserved: PBS + 0.5 mM EDTA.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| 0.5 M EDTA (pH 8.0) | 500 µL | 0.5 mM |
| 1X PBS (Ca²⁺/Mg²⁺-free) | to final volume |
Steps
- Add EDTA to Ca²⁺/Mg²⁺-free PBS.
- Filter-sterilise through 0.22 µm. Store at room temperature or 4 °C.
- Incubate cells at 37 °C for 5–15 min and tap to detach.
Good to know
- Useful before flow cytometry of trypsin-sensitive surface antigens.
⚠ Safety Handle solutions for live cells aseptically in a biosafety cabinet.
FAQ
What about cells that will not detach?
Raise EDTA to 2–5 mM or use a gentle enzyme such as accutase.
This is a widely used standard composition. Amounts depend on the hydrate form and purity of your reagents — check the formula weight (FW) on the bottle and follow your institution's safety rules and the manufacturers' instructions.