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Paraffin-embedded cell line pellet preparation

Protocol created by Lorena Maestre - Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas

  1. Grow the cell line (at least 1x108 cells are needed).
  2. Before preparing the pellet verify the status of the cell culture.
  3. Wash your pellet 2 times with PBS (5 minutes 1000prm).
  4. After the last wash resuspend you pellet in 2ml PBS and place it in a 2ml eppendorf.
  5. Centrifuge 5 min a 1300rpm.
  6. Resuspend the pellet in 2ml of formalin (10%) and mix gently (keep it in movement at room temperature during 1 hour).
  7. Centrifuge the suspension at max revolution for 5 minutes.
  8. Discard the formalin and collect the pellet using a metal spatula.
  9. Place the pellet in a special cassette. Special cassette
  10. Placed the cassette in a tissue processor.

Matherial and reagents: