Hybridoma Cell Line Medium
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1. Transfer 150 mL of RPMI (Invitrogen 21870-100) from a 500-mL bottle. (Save the remaining 350 mL for Step 3.)
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2. Add the following to the 150 mL of RPMI:
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58 mL of fetal calf serum (Gibco 10437-028), heat-inactivated for 30 min at 55°C
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5.8 mL of 100× penicillin/streptomycin (Gibco 15140-122)
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5.8 mL of 100 mm sodium pyruvate (Gibco 11360-070)
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5.8 mL of 200 mm l-glutamine (Gibco 25030-081)
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0.5 mL of 0.1% β-mercaptoethanol
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3. Filter to sterilize and add the solution back to the original bottle containing 350 mL of RPMI. Store at 4°C.
- © 2013 Cold Spring Harbor Laboratory Press
