Trypin is a serine hydrolase that can hydrolyze proteins between cells by cutting carboxyl groups in lysine and arginine residues in polypeptide chains, destroying cell connections, and thus dispersing tissues or adherent cells into single cells. The activity of trypsin in dispersing cells is related to the characteristics of tissues or cells, trypsin concentration, temperature, and action time. At pH 8.0 and 37°C, trypsin has the strongest action ability. Therefore, when using trypsin, the concentration, temperature, and time should be controlled to avoid excessive digestion and cell damage.
The working concentration of trypsin is generally 0.25%, while semi-adherent cells or cells sensitive to trypsin often use low concentrations (0.05%) of trypsin for cell digestion. Since EDTA can chelate Ca2+ and Mg2+, thereby destroying cell connections and promoting cell dissociation, a certain amount of EDTA is often added to the trypsin solution to enhance the dissociation effect.
Precautions
1. Since different tissues or cells react differently to the action of pancreatic enzymes, the operator should determine the optimal digestion time according to the actual situation; the digestion time should not be too long, otherwise it will affect the cell adhesion and growth;
2. When using this product, pay attention to aseptic operation to avoid contamination;
3. It should not be placed at room temperature or stored at 4℃ for a long time;
4. Thaw at 2℃～8℃, shake well before use, avoid repeated freezing and thawing, and it is recommended to freeze in small quantities.
5. Since the NaHCO3 content of D-Hank's balanced salt solution is low (0.35g/L), this type of trypsin solution cannot be used in a 5% CO2 environment. If placed in a CO2 culture phase, the solution will quickly become acidic, so be careful when using it;
6. This product is only used for scientific research or further research, not for diagnosis and treatment.
