Product Description/Introduction
Trypsin is a serine hydrolase that cuts off the carboxyl ends of lysine and arginine residues in polypeptide chains. Trypsin digestion is used in tissue cell extraction, culture, and in vitro cell culture to hydrolyse intercellular proteins and to dissociate tissues or cells into individual cells. Trypsin is most effectively digested at 37°C , pH 8.0. The common working concentration of trypsin is 0.25%. EDTA can chelate calcium and magnesium ions and the use of EDTA in trypsin solutions can accelerate the hydrolysis of intercellular linkages and enhance digestion. Phenol Red is a pH indicator that is purplish-red when the solution is alkaline, orange when it is acidic and pink when it is nearly neutral.
This product contains 0.25% trypsin and 0.9 mM EDTA with phenol red indicator, the solvent is PBS buffer, and is sterilized by 0.1 μm filtration.
Storage and Shipping Conditions
Ship with wet ice; Store at -20℃, valid for 12 months. Avoid freezing and thawing repeatedly.
Product Protocol/Procedures
Adherent cells
1. Aspirate the cell culture medium and wash the cells with sterile PBS, D-Hank’s solution or serum-free culture medium to remove the remaining serum.
2. Add an appropriate amount of this product to cover the cells, and let it stand for digestion for 1-5 min at room temperature.
3. Under the microscope, when the cells appear obvious contraction, it is easier to blow the cells down with the pipette. Aspirate the trypsin cell digestion solution, add the cell culture medium containing serum and gently blow down the cells to make cell suspension, which can be directly subcultured. If it is difficult to remove cells from the bottom of the plate when gently blown with a pipette, indicating insufficient digestion time, re-digest with the addition of trypsin cell digestion solution. Do not overdigest.
Note
1. Due to the different properties of tissues or cells, the experimenter should determine the optimal digestion time according to the specific situation; The digestion time should not be too long, otherwise the cell adhesion and growth will be affected.
2. This product does not contain bacteriostatic agent. Special attention should be paid to aseptic operation during use to avoid contamination of digestive juices by microorganisms.
3. This product should not be stored at 4℃ for a long time, avoid repeated freezing and thawing. It is suggested to divide into small portions and store at -20℃.
4. For the selection of EDTA and phenol red in trypsin, the trypsin digestion solution containing EDTA can be selected for general tumor cells according to experimental needs. For more sensitive and vulnerable cells, such as primary cells and easily digestible cells, choose a trypsin digestion solution without EDTA. If the cells are particularly sensitive, choose a trypsin digestion solution with a lower trypsin concentration. For apoptosis detection by flow cytometry, trypsin digestion solution without EDTA should be used.
5. This product contains phosphate system, the solution is yellow after cryopreservation at low temperature. This is due to the temperature caused by the pH value of the solution changes, in the normal thawing and rewarming that will return to red, does not affect the performance of the product. It is possible that for some reason (e.g., shipping on dry ice) the pH of the solution may drop slightly, thus changing the color of the solution from red to light yellow. This light yellow solution can still be used, or it can also be used after fine-tuning the pH with a sterile 2 M NaOH solution.
6. For your safety and health, please wear safety glasses, gloves, or protective clothing.
