Preparation and Shipping of Frozen and Live Hybridoma Cells for Antibody Production
GenScript requires two vials of frozen cells or one flask of live cells for antibody production by ascites or in vitro cell culture. Instructions for the preparation and shipping of hybridoma cells for antibody production are as follows:
Preparation and Shipping of Frozen Cells:
Only cryo-preserve rapidly growing and dividing hybridomas (log phase hybridomas) that show more than 90% viability. Under sterile conditions, harvest the log-phase hybridoma cells in a centrifuge tube and perform a cell count using a hemacytometer and trypan blue. Centrifuge the cells at 200 g (or 1,000 rpm) for five minutes at room temperature. Carefully remove the medium and re-suspend the cell pellet in freezing medium (DMEM with 20% fetal bovine serum and 10% DMSO) to final cell density of 5-10 x106 cells per ml and aliquot into cryovials at 1.0 ml/vial. Place the cryovials in cryo freezing containers or other suitable containers and place in freezer overnight at -80°C. The next day, transfer the cryo-vials to a liquid nitrogen tank. Alternatively, they can be shipped overnight if packed in a sufficient amount dry ice. Wear gloves and a facemask when working with liquid nitrogen tanks or frozen cryo-vials. If possible, send duplicate samples of each cell line.
Preparation and Shipping of Live Cells:
Only ship rapidly growing and dividing hybridomas (log phase hybridomas) that show more than 90% viability. Grow the cells to log-phase in a T-25 flask and perform a cell count using a hemacytometer and trypan blue.If the viability is less than 90%, pass the cells and continue to culture until desired cell condition is reached. Fill the culture flask with fresh culture medium,cap the flask tightly, and seal with parafilm. Vented cap flasks are not suitable for this purpose. Either use culture flasks with regular caps for cell culture or transfer the cells into regular cap flasks for shipping. Place the flask in a plastic bag and ship overnight in an Insulated Shipping Container.
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).
Our customer service representatives are available 24 hours a day, Monday through Friday to assist you.
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Email for Quotation and Ordering: order@genscript.de |
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Ordering by Telephone: 001-732-885-9188 (USA)
Toll Free Tel. No.: 0-800-0006-320 (Germany) 0800-369-254 (Switzerland) |
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Fax: 001-732-210-0262 001-732-885-5878 |
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