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drying after antigen retrieval? - (Nov/17/2008 )

Help! After antigen retrieval using citratebuffer (usually 10 minutes at 95, then slowly let cool to RT in the buffer) my student took the sections out of the buffer after 10 min at 95 and let them dry!
Is it worth it to proceed with the staining? What would be the effect of the sudden temperature change?


If I were you I would not continue the technique with those slides... The major problem I see is the fact that he let the slides dry... You may observe intense non-specific binding if you proceed... However, if you do not have more material you can give a try...


Sorry, but those slide could be thrown away. Drying of sample will give false positives results. IHC are a too timing consuming and expensive protocol to follow the protocol and at the end those results can't be use. So begin with fresh material and tell the person to not let dry out the slide until dehydrate and mounting step. Also check the time of the antigen retrieval it looks too little time usually is 20-45 min.


I would also not use them, they can be stored in PBS for a couple of days after antigen retrieval, but not dried.