Trypsin Neutralization Solution (TNS) is a sterile, phosphate and HEPES-buffered saline solution. It contains 10% fetal bovine serum as a trypsin inhibitor and cell protect agents. The product is calcium- and magnesium-free and has a pH of 7.4 at room temperature.
Incubating cells with too high a trypsin concentration for too long a time period will damage cell membranes and kill the cells. If unsure about the concentration of trypsin 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 trypsin 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 trypsin/DTA solution into culture vessel to completely cover the
monolayer of cells and place in 37°C incubator for approximately 2 minutes or until 80% of cells are rounded up (monitored with microscope).
3) Remove the trypsin/EDTA solution by aspiration and return closed culture vessel to
incubator for 1 minute.
4) Add TNS or medium containing serum to the cells as soon as possible to inhibit further tryptic 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.
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