Step I: Cell line submission, recovery, and expansion. 1-2 weeks
- Two vials of frozen hybridoma cells or one flask of live cells (> 2x106) in culture medium
- Formula of original culture medium
- Name of the species from which the cell line was derived and production history, if available
- Upon receipt, the cell line will be expanded to obtain enough cells for mouse inoculation.
Step II: Inoculation. 2-3 weeks
- The mice are primed with 0.5ml/mouse of Pristane for two weeks
- Peritoneal cavity injection of the primed mice with 0.5-1 x 106 hybridoma cells each
- Ascites production in 7-10 days
Step III: Harvest. 1 week
- Ascites harvested using aseptic techniques
- Cellular and lipid materials removed by centrifugation
- Clarified ascites aliquoted into sterile containers
- Aliquots labeled and immediately frozen
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