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Unmasking using a Pressure Cooker

A HIER/ HTAU protocol using a pressure cooker

Unmasking in a Pressure Cooker

The following protocol is for use with a pressure cooker (solution temperature ≈ 120°).

1. Cut and mount sections on slides treated with a tissue adhesive. Position slides into metal staining racks (do not place slides close together; uneven staining may occur).

 2. Deparaffinize sections and rehydrate to distilled water.

3. Add around 1600 ml unmasking solution to the pressure cooker

5. Cover, but do not lock lid, and bring solution to a boil.

6. Place slides in pressure cooker ensuring slides are well immersed in unmasking solution. Lock lid.

7. Start timing once the cooker has pressurized, start timing.

 8. After one minute, remove the pressure cooker from heat source and run under cold water. DO NOT OPEN LID UNTIL THE INTERNAL PRESSURE HAS BEEN COMPLETELY REDUCED.

9. Open lid, remove slides and place immediately into a tap water bath. DO NOT LET SECTIONS DRY OUT. Cooler water can be added to the pan to cool sections down before transferring – do not remove very hot sections from solution.

10. Wash sections in PBS buffer (pH 7.5) for 5 minutes.

Optimize treatment time for individual use.

** Follow the manufacturer’s instructions and safety recommendations for use.

Commercially available high & low pH antigen unmasking solutions are suitable for this method. To get a quote for the products or to place an order, please email