[Histonet] IHC on paraffin embedded skin tissue!

[Jennifer Leigh]

       Histonetters-
 
           I have a question about mouse skin samples----an IHC experiment has gone wrong and I am not sure why. I have a set of paraffin embedded mouse skin samples from an inflammatory model that were stained for F4/80. The end result is no color development after the addition of DAB. I use the DAB tablets from Sigma using the TBS buffer to dissolve the tablet. There is no color development, not even non-specific staining. Any ideas what the problem might be?????  I also performed a Proteinase K antigen retrevial step (using the ready made solution from Dako) for 15 minutest at 37 C as I was instructed to do. Thank you in advance for any insight!
 
Jennifer Oskins

Jennifer L. Oskins
"Until one has loved an animal, part of their soul remains unawakened....."