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The protocols for heat-induced epitope retrieval and enzymatic epitope retrieval.
Heat-Induced Epitope Retrieval
Decloaker
- Add 500ml distilled water to the pan of the Decloaker™.
- Place a full rack of slides in a Tissue Tek™ container containing 250ml of retrieval solution.
- Decloak for 5 minutes.
- Depressurize for 10 minutes.
- Remove pan top and cool for 10 minutes.
- Rinse in distilled water for 3 minutes. Repeat twice.
Equipment information: Decloaker
Microwave (Panasonic Model No. NN-S744WF 1200 Watts)
- Place a full rack of slides in a Tissue Tek™ container containing 250ml of retrieval solution.
- Microwave for 5 minutes at level 5.
- Cool for 1 minute (Add 50ml of retrieval solution to the container, if necessary).
- Microwave again for 5 minutes at level 5.
- Remove the slides from the microwave oven and cool 20 minutes at room temperature.
- Rinse slides in distilled water for 2 minutes. Repeat twice.
Equipment information: Microwave
Steamer
- Place two containers in the steamer basket.
- Fill container 1 with 250ml of 1X Automation Buffer.
- Fill container 2 with 250ml of retrieval solution.
- Turn steamer dial to 35 minutes.
- Place slides in container 1 for 5 minutes.
- Remove slides, drain and transfer to container 2 for 30 minutes.
- Remove slides from the steamer and cool at room temperature for 20 minutes.
- Rinse slides in distilled water for 1 minute to stop the reaction.
- Incubate slides in 1X Automation Buffer for 5 minutes.
- Proceed with stain.
Equipment information: Steamer