Enzyme-Free Cell Dissociation Solution Protocol

1. All reagents should be warmed to 37°C.
2. Withdraw growth media from cells.
3. Rinse cells with Hank’s BSS (w/o Ca & Mg) or PBS (w/o Ca & Mg). Use approximately 5 mL in a 75 cm² flask; gently rock the flask (or plate) back and forth, allowing the solution to bathe the cells for 30 to 60 seconds. Withdraw the rinse solution and discard.
4. Add approximately 5 mL of Enzyme-Free Cell Dissociation Solution to the 75cm² flask (or 100 mm plate). Gently rock the vessel back and forth for 1 to 2 minutes, at room temperature. Withdraw the solution.
5. Firmly tap vessel against the palm of hand to dislodge cells. If cells do not dislodge quickly, allow vessel to sit at room temperature for an additional 2 to 5 minutes and again tap vessel against palm of hand. This step may need to be repeated for strongly adherent cells. After the cells are visibly detached add complete growth media. Resuspend cells and pipette repeatedly to break up any clumps that may be present with certain cell types. The cells are now ready to be used in your experimental procedure.