Immunoperoxidase Protocol for Dako Autostainer
- Cut tissue sections at 3-5 microns and mount on appropriate control box slides.
- Bake slides for 30 minutes in 70°C - 90°C oven.
- Label slides with necessary information including antibody.
- Deparaffinize slides in 3 changes of xylene 5 minutes each.
- Bring slides through 2 changes of absolute alcohol and 1 change of 95% alcohol 2 minutes each.
- Rinse in 2 changes of distilled waters. Then place in buffer solution.
- Perform any digestion or antigen retrieval protocols now if necessary.
- Set up DAKO autostainer with all reagents and antibodies necessary.
- Load slides and start machine.
- When completed, remove slides and wash in running tap water 2-5 minutes.
- Dehydrate slides
Rinse 15-20 dips in
- 2 changes 95% ETOH
- 2 changes ABS ETOH
- 3 changes Xylene
- Coverslip on automatic coverslipper.
