Can I perform alternative antigen retrieval methods than those provided in the SignalStar™ protocol?

For optimal results, we don’t recommend deviating from SignalStar protocols. However, if you don’t have access to the antigen retrieval method detailed in the protocol, you may try using a microwave, which has shown to result in reduced fluorescent signal. We don’t recommend using a microwave when detecting low abundance markers, and can’t guarantee your result with this alternative method.

To perform antigen retrieval using a microwave:  

1. Prepare 250 mL of 1X EDTA unmasking solution by diluting 25 mL of SignalStain® EDTA Unmasking Solution (10X) (#14747) with 225 mL of dH₂O.
2. Submerse slides in a container of 1X EDTA unmasking solution and heat in a microwave until boiling is initiated (~2.5 minutes at power level 10). 
3. Follow with 15 minutes at a sub-boiling temperature (95°-98°C). In most microwaves, this equates to 8 minutes at power level 3, then 7 minutes at power level 2. 
4. Remove the container of slides from the microwave and place under a dH₂O faucet. Add water directly to the slide container for 5 minutes until all EDTA is replaced by dH₂O. No cooling is necessary.

Last updated: September 18, 2024