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Formulation of media for Hybridoma cells

This media is for maintaining mouse hybridoma cells (established monoclonal lines or newly expanded clones).

Base media: Dulbecco’s Modification of Eagle’s Medium: 1X, with 4.5 gm/L Glucose, with L-Glutamine, with Sodium Pyruvate (Mediatech #45000-304)

Supplemented with:

  • 10% Fetal Bovine Serum (50 ml) (PAA, ###)
  • 50 µM 2-Mercaptoethanol (0.5 ml) (make 50 mM stock in PBS or sterile H2O, filter sterilize)
  • 10 mM HEPES (5 ml) (1:100 of 1M stock, Mediatech #15630-080)
  • MEM Non-Essential Amino Acids (5 ml) (1:100 of 100x stock, Mediatech #45000-700)
  • Penicillin/Streptomycin (5 ml) (100X aliquots in media prep freezer, Mediatech #45000-652)

Mouse monoclonal lines tested:

  • Hyb9901
  • Y3
  • c127
  • c136

(Mediatech products distributed through VWR)

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