Tissue digestive enzyme

         This product contains various proteases such as collagenase commonly used in enzymatic tissue digestion, which can specifically degrade extracellular collagen and dissociate the network structure of collagen and fibronectin without damaging the integrity of the cell surface structure. Using this product for tissue digestion can loosen the bridging structure between cells in the tissue, transforming the tissue from blocky to flocculent. After mechanical blowing, a small number of cell clusters and a large number of individual cell suspensions are produced. The prepared cell suspension can be used for primary culture and flow cytometry for cell biology research such as apoptosis, proliferation, cell cycle, etc. Scope of application: Various normal and tumor tissues, from which primary cells, including epithelial cells, stromal cells, fibroblasts, etc., are isolated.

Instructions for use:

(1) Take out the digestive enzymes from the -20 ℃ refrigerator in advance and melt them at 2-8 ℃;

(2) Dilute tissue digestive enzymes in a 1:1 ratio using a diluent of digestive enzymes to prepare a working concentration;

(3) Transfer the tissue to a culture dish, add 3 mL of tissue cleaning solution to rinse the tissue, repeat 2-3 times;

(4) Mechanically cut the tissue into 1-2 mm3 sizes under sterile conditions and transfer it to a 15 mL sterile centrifuge tube;

(5) Add 5 mL of tissue cleaning solution and centrifuge at 1500 rpm for 3 minutes at room temperature;

(6) Abandon the supernatant, add 12mL of working concentration digestive enzyme, label the sample name and number, seal it, and place it in a constant temperature oscillation incubator for digestion. Set the parameters at 37 ℃, 200-300 rpm (set the speed according to the sample texture), and let the tissue digest in a flocculent form without obvious hard blocks visible to the naked eye;

(7) Take out the digested sample and centrifuge at 1500 rpm for 3 minutes at room temperature.

(8) After discarding the supernatant, add 6mL of serum containing medium for digestion termination, and centrifuge at 1500 rpm for 3 minutes at room temperature.

(9) Discard the supernatant and add the diluent for use.

Matters needing attention

1. This product needs to be melted at 2-8 ℃ before use, and should be used within one week after melting.

Storage conditions: -20 ℃, stored in dark for 6 months, valid.

Note: For scientific research purposes only.