Freezing S2 Cells

  1. Spin ~3X107 cells @ 500xg for 5min.

  2. Save 1.5 ml conditioned media

  3. Mix w/ 1.5 ml fresh media

  4. Add 225 ml DMSO (7.5%)

  5. Sterile filter 3.0 ml media

  6. Resuspend cells in sterile freezing media

  7. Aliquot 1.5 ml in 2 ml cryovials

  8. Freeze in -80°C freezer overnight (in foam box to slow cooling)

  9. Store long-term in liquid N2