Trypsin is a serine hydrolase that can cut off the carboxyl ends of lysine and arginine residues in polypeptide chains. Trypsin digestion solution is used in the extraction and culture of tissue cells and in vitro cell culture to hydrolyze intercellular proteins and separate tissue or cells into single cells. Trypsin digestion was the strongest at 37℃ pH 8.0. The usual working concentration of trypsin is 0.25%. EDTA can chelate calcium and magnesium ions. Using EDTA in trypsin solution can accelerate the hydrolysis of intercellular conjunctions and enhance digestion. Phenol red is a pH indicator, the solution is more basic when purplish red, more acidic when orange red, nearly neutral when pink.

 

 

 

This product is 0.25% trypsin digestive solution, containing 0.25% trypsin, soluble in Hank's buffer, edTA-free, containing phenol red indicator, filtered by 0.22 μm for bactericide.

 

 

 

 

 

 

 

Storage and transportation

 

 

 

Dry ice transportation; Stored at -20℃, valid for 12 months. Avoid repeated freezing and thawing.

 

 

 

 

 

 

 

Instructions for

 

 

 

Adherent cells

 

 

 

1. Suck the cell culture medium and wash the cells with sterile PBS, Hanks solution or serum-free culture medium to remove the residual serum.

 

 

 

2. Add an appropriate amount of this product to cover cells and let stand at room temperature for digestion for 1-5 min.

 

 

 

3. Under the microscope, when the cells contracted significantly and it was easier to blow the cells down by gently blowing them with a pipette gun, the digestive juices of pancreatic cells were sucked out, cell culture medium containing serum was added, and the cells were gently blown down to make cell suspension, which could be directly subcultured. If it is difficult to make the cells fall off from the bottom of the plate when gently blowing with a pipette gun, indicating insufficient digestion time, trypsin cell digestive fluid can be added for re-digestion. Avoid excessive digestion.

 

 

 

 

 

 

 

Matters needing attention

 

 

 

1. Due to different tissue or cell properties, the experimenter should determine the best digestion time according to the specific situation; Digestion of cells should not take too long, otherwise it will affect cell adherence and growth.

 

 

 

2. This product does not contain bacteriostatic agent. Special attention should be paid to aseptic operation during use to avoid microbial contamination of digestive juices.

 

 

 

3. The product should not be stored at 4℃ for a long time to avoid repeated freeze-thaw. It is recommended to store in small portions at -20℃.

 

 

 

4. For the selection of EDTA and phenol red in pancreatin, trypsin digestive solution containing EDTA can be selected for general tumor cells according to experimental needs. For more sensitive and fragile cells, such as primary cells and easily digestible cells, choose trypsin digestion solution without EDTA. If cells are particularly sensitive, choose digestive solution with lower trypsin concentration. If it is necessary to detect apoptosis by flow cytometry, trypsin digestion solution without EDTA is used.

 

 

 

5. Due to the low NaHCO3 content in Hank's solution, it is not recommended to use this product for digestion in a CO2 incubator at 37 ° C, the solution will quickly become acidic if the CO2 phase is added.

 

 

 

6. For your safety and health, please wear lab coat and disposable gloves.

 

 

 

 

 

 

 

Products for scientific research purposes only, not for clinical diagnosis!