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Heat antigen retrieval

Material:
Hot plate
2000ml glass beaker
250ml plastic container
slides rack
slides container
Thermometer
Buffer: Bulls Eye (BULL1000L2JX BioCare) or 0.01M sodium citrate buffer, pH 6.0

Procedure:

1. Fill glass beaker with 1000ml ddH₂O.
2. Put a thermometer in the beaker.
3. Put beaker on hot plate and turn on the hotplate.
4. Check water temperature until it is stable at the desire temperature (95-99°C or 102-104°C).
5. Use a 250ml glass bottle and fill with 190ml ddH₂O. Warm 30 minutes before use it.
6. Deparaffinize the slides and keep slides in the container with ddH₂O.
7. Take bottle from water bath and add 10ml 20X bulls eye solution to the bottle. Shack well.
8. Empty ddH₂O in slides container and pour 1X bulls eye solution to slides container.
9. Put Container back to water bath as soon as possible.
10. Heat for 20 min.
11. Wash slide with ddH₂O 2-3 times Slides are ready for immunostaining.

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