Protocols

Trypsin Digestion of Formalin-Fixed, Paraffin-Embedded Tissues Prior to Immunohistochemistry

1. Preheat the following to 37°C using a water bath:
   200ml of TBS
   200ml of distilled water.
2. Dissolve 0.2g Trypsin 250 and 0.2g Calcium Chloride in the 200ml of TBS.
3. Once the trypsin solution is at 37°C, pH to 7.8 with 1M sodium hydroxide.
4. Place rehydrated paraffin sections in the distilled water to preheat the sections to 37°C for a minimum of 5 minutes.
5. Incubate sections in trypsin solution at 37°C. The time required will depend on the antibody and tissue, however, 30 minutes is usually sufficient.
6. Rinse sections in running tap water.
7. Proceed with immunohistochemistry protocol.

NB. Trypsin containing chymotrypsin should always be used. The enzyme activities can vary from a supplier and between batches. Such Variations may affect the incubation time required.

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