1. Cut and mount 3-5 micron formalin-fixed paraffin-embedded tissues on positive charged
slides such as Bio SB Hydrophilic Plus Slides (BSB 7028).
2. Air dry for 2 hours at 58° C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as
ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).

5. Any of three heating methods may be used:
a. TintoRetriever Pressure Cooker or Equivalent
Place tissues/slides in a staining dish or coplin jar containing the ImmunoDNA Retriever
with Citrate or EDTA, and place in the pressure cooker. Add 1-2 inches of distilled water to
the pressure cooker and turn heat to high. Incubate for 15 minutes. Open and immediately
transfer slides to room temperature.
b. TintoRetriever PT Module or Water Bath Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA
Retriever with Citrate or EDTA at 95°-99° C. Incubate for 30-60 minutes.
c. Conventional Steamer Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA
Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room
temperature and let stand for 15-20 minutes.
7. Wash slides with IHC wash buffer or DI water.
8. Continue IHC staining protocol.