Water Bath Unmasking Protocol
Using a water bath at temperatures just below boiling can be a convenient method for many laboratories. Temperatures as low as 90°C can be used in conjunction with longer incubation times; the time can be shortened at 95-98°C.
Use of a water bath below boiling should mean that loss of solution through evaporation is not an issue, but tissue sections should never be allowed to emerge from hot solution.
- Cut & mount sections on coated slides - VECTABOND, Poly L-Lysine or APES coatings are recommended
- Deparaffinize sections and rehydrate to distilled water
- Fill a coplin jar with unmasking solution and pre-warm to the desired temperature in a temperature controlled water bath.
- Add dewaxed and rehydrated sections to a slide rack and completely immerse slides in the coplin jar.
- Incubate for 20-40 minutes. If sections require different unmasking times, they can be added throughout this incubation, ensuring timings mean all slides finish together.
- Protecting yourself from the steam, remove coplin jar with slides from the water bath and allow to cool to room temperature. DO NOT REMOVE SLIDES UNITL COOLED TO ROOM TEMPERATURE.
- Wash in buffer.
Commercially available high & low pH antigen unmasking solutions are suitable for this method. To get a quote for the products or to place an order, please email Sales@2BScientific.com.