EDTA-Free Cell Dissociation Buffer (100mL)
Features: Cell Dissociation Buffer, Enzyme-free, Animal Origin-Free, Human Origin-Free, Room Temperature Stable, Phenol Red-free
This product is sterile (membrane-filtered) and ready to use, isotonic, and enzyme-free solution of salts, chelating agents, and cell-conditioning agents in calcium-free and magnesium-free Solution.
1. Warm all reagents to 37°C prior to use.
2. Remove growth medium from cells.
3. Thoroughly rinse cell monolayers with 5 ml Ca++- and Mg++-free PBS per T75 flask or 100 mm dish. Gently rock the flask (or dish), allowing the solution to bathe the cells for 30 to 60 seconds at room temperature. Aspirate the rinse solution and discard.
4. Repeat step 3.
5. Add approximately 5 ml of Cell Dissociation Buffer per T75 flask or 100 mm dish and gently bathe cells by rocking at room temperature for ~1 minute. You may check for dissociation under the microscope. Aspirate solution and discard.
6. Add approximately 5 ml of Cell Dissociation Buffer per T75 flask or 100 mm dish and culture in 37°C incubator for 3-5 minutes. Take out the cells from the incubator. Firmly tap flask or dish against palm of hand to dislodge cells. If cells do not detach quickly, allow to sit at room temperature for another 2 to 5 minutes and tap flask against palm of hand again. This may be repeated for cells that are more strongly adherent (with 5 more mls of dissociation buffer). After the cells are visibly detached add at least 5 ml of complete growth medium. Resuspend cells in growth medium.