Product Description
Enzyme-free Cell Dissociation Solution (ECDS) is a sterile, phosphate and HEPES-buffered saline solution. It contains 5mM EDTA, 1 mM sodium pyruvate and 10 mM HEPES. This product has a pH of 7.4 at room temperature.

Product Use
ECDS is used to detach adherent primary cells from a culture surface. It is for research use only. Not for use in animals, humans, or diagnostic procedures.

Storage
Store the Enzyme-free Cell Dissociation Solution at -20°C. Once thawed, the product may be stored at 4°C for up to one month.

Shipping
Dry ice.

Procedure
Incubating cells with too high a EDTA concentration for too long a time period will damage cell membranes and kill the cells. If unsure about the concentration of EDTA to use, use a low concentration. The time required to remove cells from the culture surface is dependent on cell type, population density, and serum concentration in the growth medium. The time of EDTA exposure should be kept to a minimum.
1) Remove medium from culture vessel by aspiration and wash the monolayer with Ca+2 and Mg+2-free salt solution to remove all traces of serum. Remove salt solution by aspiration.
2) Dispense enough Enzyme-free Cell Dissociation Solution into culture vessel to completely cover the monolayer of cells and place in 37°C incubator until 80% of cells are rounded up (monitored with microscope).
3) Remove the Enzyme-free Cell Dissociation Solution by aspiration and return closed culture vessel to incubator for 2 minutes.
4) Add medium containing 5 – 10% serum to the cells as soon as possible to inhibit further dissociation activity which may damage cells.
5) Cells can be harvested by gently pipetting the cell suspension. Further dilution can be made, if required, for cell counts and/or subculturing.
Caution: If handled improperly, some components of this product may present a health hazard. Take appropriate precautions when handling this product, including the wearing of protective clothing and eyewear. Dispose of properly.