Details
Molecular formula: C6H15O12P3
Molecular weight: 24000
The trypsin-EDTA solution (Trypsin-EDTA Solution) contains 0.25% trypsin and 0.02% EDTA (0.53mM), dissolved in calcium and magnesium-free balanced salt solution, sterilized by filtration, and can be used directly Digestion of cultured cells and tissues. This product has the characteristics of convenience, speed, stability and safety, and good cell status.
Product Description:
In the in vitro culture of tissue cells and the dispersion of tissue cells in primary cell culture (preparation of tissue blocks into a single cell suspension) and passage cell culture, the digestion and dispersion of adherent growth cells should use tissue cell digestion solution. Commonly used digestive solutions are trypsin, EDTA, etc., whose main function is to hydrolyze intercellular proteins (such as extracellular matrix), disperse tissues or adherent cells into single cells, and make cell suspensions for further experiments.
Storage: Store at -20°C, valid for 12 months.
Instructions:
1. Digestion of Adherent Cells:
a) Aspirate the culture medium, and wash the cells once with sterile PBS, Hanks solution or serum-free medium to remove residual serum.
b) Add a small amount of trypsin digestion solution to cover the cells and let stand at room temperature for 1-2 minutes. Different cells have different digestion times, and the digestion time can be extended appropriately for cells with firm adherence.
c) Observing under a microscope, the cells shrink obviously, and the shape of the cells changes significantly when observed at the bottom of the culture vessel with the naked eye; or blowing the cells with a gun shows that the cells can just be blown off. At this point the digestive juices are aspirated. Add serum-containing cell culture medium, blow down the cells, and then directly use them in subsequent experiments.
d) If insufficient digestion is found, trypsin digestion solution can be added to re-digest.
e) If it is found that the digestion time of the cells is too long, some of the cells have fallen off the bottom of the culture vessel before pipetting the cells, and all the cells are directly blown off with trypsin cell culture medium. Centrifuge at 1000-2000g for 1 minute to pellet the cells, try to remove the trypsin cell digestion solution, add the complete culture medium containing serum to resuspend the cells, and then use it for subsequent experiments.
2. Digestion of tissue:
The time required for digestion of different tissues varies greatly, and it is generally appropriate to fully break up the tissues after digestion.
Precautions:
1. Due to the different properties of tissues or cells, the experimenter should determine the optimal digestion time according to the specific situation; the time for digesting cells should not be too long, otherwise it will affect the cell adhesion and growth;
2. This product does not contain bacteriostatic agents, special attention should be paid to aseptic operation during use to avoid microbial contamination of the digestive juice;
3. It is not suitable for long-term storage at 4°C, and repeated freezing and thawing should not be avoided. When using in small quantities, it is recommended to pack and freeze;
4. For your safety and health, please wear a lab coat and disposable gloves for operation.
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