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Instructions on Preparation and Shipping of Frozen Hybridoma Cells to GenScript for Antibody Production

GenScript requires two vials of frozen cells for antibody production by ascites or in vitro cell culture. Please follow the instructions below for the preparation and shipping of hybridoma cells for antibody production. Please let us know if the hybridomas have Balb/c or other significant genetic backgrounds. We will also need to know the medium in which the hybridomas were cultured (DMEM or RPMI). Please provide a completed Hybridoma Cell Line or Antibody Information Form with your frozen cells.

Please download the Hybridoma Cell Line or Antibody Information Form

Preparation and Shipping of Frozen Cells:

Only cryo-preserve rapidly growing and dividing hybridomas (log phase hybridomas) that show more than 90% viability. If possible, send duplicate samples of each cell line.

1. Under sterile conditions, harvest log-phase hybridoma cells in a centrifuge tube and perform a cell count using a hemacytometer and trypan blue.
2. Centrifuge cells in a centrifuge tube at 200 g (or 1,000 rpm) for five minutes at room temperature.
3. Carefully remove the medium and re-suspend the cell pellet in freezing medium (DMEM with 90% fetal bovine serum and 10% DMSO) to a final cell density of 5-10 x10⁶ cells per ml.
4. Aliquot into cryovials at 1.0 ml/vial.
5. Place the cryovials in cryo freezing containers or other suitable containers and place in freezer overnight at -80°C.
6. The next day, transfer the cryo-vials to a liquid nitrogen tank. Alternatively, the frozen cells can be shipped overnight if packed in a sufficient amount of dry ice. Wear gloves and a facemask when working with liquid nitrogen tanks or frozen cryo-vials.