Protocol for microwave preparation of paraffin tissue embedded sections

1. Deparafinize in xylene for 10 minutes. Repeat twice for a total of three treatments
2. Hydrate sections through graded alcohols
3. Rinse three times with distilled water
4. Incubate for 10 minutes with 0.1% H₂O₂ in PBS to quench endogenous peroxidase activity
5. Rinse four times with PBS
6. Place slides in a staining dish and fill container with 10mM citrate buffer (do not seal)
7. Place dish in microwave. Either set the probe to 190^oF or see the following reference to calibrate your microwave: Tacha, DE, Chen, T, Modified Antigen retrieval procedure: calibration technique for microwave ovens. Histotechnology 17(4): 365, 1994
8. After correct temperature has been reached, gently mix the 10mM citrate buffer with a disposable pipet
9. Hold temperature in the microwave for approximately 5 minutes
10. Take the slides out and allow to equilibrate to room temperature
11. Rinse slides three times with PBS
12. Slides are ready for staining