Hybridoma Cell Specific Serum/Protein-Free Non-gradient Cell Freezing Medium

Serum-free protein-free non-programmed cell cryopreserved solution for hybridoma cells is a ready-to-use non-programmed cell cryopreserved solution with clear chemical composition, no serum, no protein and containing DMSO. This product is chemically defined and serum-free, which can effectively avoid the unstable results of serum batch instability and unknown composition on cell freezing. Serum-free and protein-free, the potential effects of certain proteins on some specific cells can be avoided and applied to serum-free culture cells or expressed cells for cryopreservation. In addition, compared with traditional cryopreservation medium, this product take the non-gradient freezing, only need to resuspend the cells with this product and put them directly into -80℃ refrigerator, eliminating the operation of tedious gradient freezing.

This product has been continuously optimized and debugged by our technical team, and while it has the above advantages, it greatly weakens the crystallization process of water, protects cells from solute damage, and effectively improves viability and cell recovery after thawing. Special optimization is made for sensitive cell lines such as hybridoma, which can support the direct freezing of hybridoma cells without digesting the whole plate.

Ship with wet ice; Store away from light at 2-8℃, valid for 12 months.

1. Hybridoma routine cryopreservation scheme:

1.1. Hybridoma cells in good condition are collected in centrifuge tubes according to conventional methods;

1.2. Centrifuge at 800-1200rpm/min for 3-5min, remove the supernatant, and retain cell precipitation;

1.3. Adding serum-free protein-free non-programmed frozen solution for hybridoma cells, gently blow the heavy suspension cells, and the cell density is controlled within 1×106~5×106 /mL;

1.4. Transfer the above cellular freeze-storage solution to the cell freeze-storage tube in an appropriate volume (1mL/ branch is recommended);

1.5. Freeze the cell cryotubes directly in a -80°C refrigerator;

1.6. The next day transfer to the liquid nitrogen tank for long-term storage, can also be placed in the -80℃ refrigerator for short-term storage.

2. Hybridoma full plate freezing protocol during subcloning (96-well plate as an example) :

2.1. Pre-warm the serum-free protein-free non-programmed cryopreservation solution for hybridoma cells by placing it in culture at 37°C in advance;

2.2. Remove the original cell culture medium and add a certain volume of serum-free, protein-free, non-programmed cryopreservation solution for hybridoma cells (100 μL for 96-well plates);

2.3. After sealing the openings of the cell-well plate with a sealing film, the cell-well plate was directly placed flat into a -80°C refrigerator for freezing;

2.4. For recovery, thaw the cell plate in a 37°C incubator (can be placed on top of a shaker to accelerate thawing);

2.5. After thawing, aspirate the supernatant directly and add 200 μL of fresh medium for incubation; or mix the cells with a plate, transfer 30 μL of each well into a new plate and make up 200 μL of fresh medium for incubation.

1. Gently shake the reagent before use to ensure even distribution of each component.

2. Please use up the product within 3 months after opening, also can be divided and put in -20℃ for long-term storage.

3. Make sure the cells are in good condition before freezing, and make sure the corresponding markers are made on the cell cryotube. Use marker pen that are resistant to organic solvents to avoid loss of markers on the cell cryotube.

4. The product has a certain degree of viscosity, the routine frozen storage program recovery do not directly centrifuged, it is recommended to dilute and then centrifuged before the seed plate; the whole plate frozen storage in accordance with the instructions of the program steps can be carried out.

5. For your safety and health, please wear safety glasses, gloves or protective clothing.

For Research Use Only!