Product Information
|
Product Name |
Cat.No. |
Spec. |
|
0.25% Trypsin (Soluble in D-Hank's, EDTA,no Phenol Red) |
G4012-100ML |
100 mL |
Introduction
Trypsin, abbreviated as trypsin, is a serine hydrolase that cuts off the carboxyl termini of lysine and arginine residues in polypeptide chains. In the process of tissue cell extraction, culture, and in vitro cell culture, trypsin digestion solution is used to hydrolyze intercellular proteins, so that the tissue or cell can be dispersed into a single cell. The digestion ability of trypsin was the strongest at 37℃ pH 8.0. The common working concentration of trypsin is 0.25%. EDTA can chelate calcium and magnesium ions. The use of EDTA in trypsin solution can accelerate the hydrolysis of intercellular junction proteins and enhance digestion. Phenol red is a pH indicator. The solution is purplish-red when it is more basic, orange-red when it is more acidic, and pink when it is near neutral.
This product is a 0.25% trypsin digestion solution, containing 0.25% trypsin, dissolved in Hank's buffer, containing 0.9 mM EDTA, without phenol red indicator, sterilized by 0.1 μm filtration.
Storage and Handling Conditions
Transport with dry ice; Store at -20℃, valid for 12 months. Avoid repeated freezing and thawing.
Usage
Adherent cells
1. Blot off the cell culture medium and wash the cells with sterile PBS, Hanks 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 by gently blowing the cells with a pipette gun. Then the trypsin cell digestion solution is removed by suction, and the cell culture medium containing serum is added to 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 gun, 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 best digestion time according to the specific situation; The time of digesting cells 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 to avoid repeated freezing and thawing. It is suggested to be divided into small parts and stored at -20℃.
4. For the selection of EDTA and phenol red in pancreatic enzymes, 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. If it is necessary to detect apoptosis by flow cytometry, trypsin digestion solution without EDTA should be used.
6. For your safety and health, please wear a lab coat and disposable gloves when operating.
