This product is used for washing tumor or normal tissues during tissue pretreatment. It contains a combination of multiple antibiotics to exert synergistic antibacterial effects, which can effectively reduce contamination rates in primary cell culture.
Instructions for Use
1. Take the digestive enzyme out of the -20℃ refrigerator in advance and thaw it at 2–8℃.
2. Dilute Tissue Digestion Enzyme (Catalog No. O1521148) at a ratio of 1:1 with Digestive Enzyme Diluent (Catalog No. D1521149) to prepare working solution.
3. Transfer tissues into a petri dish, add 3 mL tissue washing solution to rinse tissues, and repeat rinsing 2–3 times.
4. Under aseptic conditions, mechanically cut tissues into 1–2 mm³ fragments, then transfer them into sterile 15 mL centrifuge tubes.
5. Add 5 mL tissue washing solution and centrifuge at 1500 rpm at room temperature for 3 minutes.
6. Discard the supernatant, add 12 mL working-concentration digestive enzyme. Label sample names and numbers, seal the tubes, and place them in a constant-temperature shaking incubator for digestion. Set parameters at 37℃, 200–300 rpm (adjust speed according to tissue texture). Terminate digestion when tissues become flocculent with no visible hard lumps.
7. Take out digested samples and centrifuge at 1500 rpm at room temperature for 3 minutes.
8. Discard the supernatant, add 6 mL serum-containing medium to terminate digestion, and centrifuge at 1500 rpm at room temperature for 3 minutes.
9. Discard the supernatant, resuspend cells with primary cell medium for subsequent use.
Precautions
1. Strict aseptic operation is required during use to avoid contamination.
2. Use the product within the shelf life. Ideal experimental results cannot be guaranteed after expiration.
3. For research use only, not for in vitro diagnosis.
ZJ26F0636572