[Histonet] deparaffinization won't work!


Dear Histoneters,
I am trying to do an FMRP immuno on brain specimens that were fixed and embedded
in paraffin. Unfortunately I couldn't get staining. So I did a Nissl and I don't
get staining with that as well so I am guessing my tissue isn't fully
I have tried everything from:
-Preheating the tissue on a hot plate for 2 hours.
-Spraying the sections with ParaGuard (a paraffin repellant used to remove
paraffin stains) after heating the sections.
My protocol for deparaffinization is currently:
-2 times 30 min in Xylene
-10 minutes in 100% Ethanol
-5 minutes in 95% Ethanol
-5 minutes in 70% Ehtanol
-5 minutes in 50% Ethanol
-2 times 5 minute washes in 0.05% PBST
-2 rinses in dH2O.

Then I do an antigen retrieval step with Vector antigen Retrieval kit and then
my immuno.

Any help would be greatly appreciated.
Sophia Koukoui
MBNS, Psychology
McGill University
Montreal Canada.

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