I have already stained FFPE rat kidney's using FITC with a 2 hour heat treatment in citrate buffer ph 6.0.  (the staining was great)

I am now going to restain the same rat kidney slides with an additional primary using DAB as the chromagen.  This primary also requires heat treatment.

Am I correct in my thinking that I should repeat the heat treatment again due to the fact that the epitope (binding) sites have already closed after accepting the application of the 1st primary?

Any help will be appreciated.  Thank you in advance.

Carol Ann Bobrowitz
Medical College of Wisconsin
Department of Physiology
414-456-8179
cbobrowi@mcw.edu
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