| Trypsin is a serine hydrolase that can cleave the base-side segments of lysine and arginine residues in polypeptide chains, hydrolyzing the proteins between cells and disrupting cell connections, causing tissues or adherent cells to disperse into single cells. The activity of trypsin is related to the characteristics of the tissues or cells, the concentration of trypsin, the temperature, and the duration of action. Trypsin performs best at pH 8.0 and 37°C. Therefore, it is important to control the concentration of trypsin, temperature, and exposure time to avoid excessive cell damage. Generally, the commonly used working concentration of trypsin is 0.25%, while for semi-adherent or adherent cells, the concentration is 0.05%. Since EDTA can bind Ca2+ and Mg2+, destroying cell connections and promoting cell dissociation, a certain amount of EDTA is often added to trypsin solutions to enhance the dissociation effect. |
