PET Cell Dissociation Reagent is a cell dissociation mix specially formulated for use with the "Cold Trypsin" method. When used to detach monolayer cultures of epithelial cells from the growth substratum, this reagent will yield more viable cells of monolayers grown in serum-free medium. The solution contains polyvinylpyrrolidone, EGTA and trypsin in a HBS base.
Suggested Use:
After rinsing the cell monolayers using HBS, add 3ml of ice cold PET dissociation reagent. Do not keep the flask in the incubator. Incubate at room temperature and monitor the monolayer by microscopy (preferably phase contrast). When the cells begin to separate, gently remove most of the PET (without removing the cells) and allow the flask to remain at room temperature. To determine whether the cells are ready to harvest, tap the flask (held in an upright position) firmly by hand.
When the cells float free, resuspend them in cold HBS supplemented with 5% Fetal Bovine Serum to stop the tryptic action. Mix well by pipetting the cell suspension, centrifuge and resuspend the cell pellet in the appropriate complete serum-free medium.
pH: 7.2-7.6 (5% CO2 atmosphere is recommended for culture.)
Conductivity:
As reported
Trypsin Activity: Determined using PDQTM Protease Assay.
Sterility Test:
No detection of microbial growth.
Storage and Stability:
May be stored at 4 degrees C for short-term (4-6 weeks). For long-term storage, aliquot to avoid repeated freezing and thawing and freeze at -70 degrees C. Aliquots are stable for 6 months.
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