Early Detection Research Network

Immunoperoxidase Protocol for Dako Autostainer

  1. Cut tissue sections at 3-5 microns and mount on appropriate control box slides.
  2. Bake slides for 30 minutes in 70°C - 90°C oven.
  3. Label slides with necessary information including antibody.
  4. Deparaffinize slides in 3 changes of xylene 5 minutes each.
  5. Bring slides through 2 changes of absolute alcohol and 1 change of 95% alcohol 2 minutes each.
  6. Rinse in 2 changes of distilled waters. Then place in buffer solution.
  7. Perform any digestion or antigen retrieval protocols now if necessary.
  8. Set up DAKO autostainer with all reagents and antibodies necessary.
  9. Load slides and start machine.
  10. When completed, remove slides and wash in running tap water 2-5 minutes.
  11. Dehydrate slides

    Rinse 15-20 dips in

    • 2 changes 95% ETOH
    • 2 changes ABS ETOH
    • 3 changes Xylene
  12. Coverslip on automatic coverslipper.