PROCEDURE - STEPWISE
1. Paraffin sections mounted on pretreated slides are heated in an oven for
30 minutes.
2. Deparaffinize sections using 4 changes of xylene for 5 minutes each, followed by 4 changes of 100% ethanol for 3 minutes each. For sections that do not require digestion, go directly to Step 6.
3. For sections requiring digestion, rinse in deionized water for 3 minutes.
4. Incubate sections in buffered protease VIII at 370C for 1 to 30 minutes. The standard time for digestion is 3 minutes.
5. Rinse sections in 2 changes of deionized water for 2 minutes each, then
3 changes of 100% ethanol for 3 minutes each.
6. Block endogenous peroxidase activity with 3% H202 methanol for 30 minutes.