IHC with glutaraldehyde-fixed tissue. - (Mar/06/2015 )

I'm exploring the possibility of performing fluorescent immunohistochemistry on paraffin embedded tissue. My problem is that the tissue was fixed with 2% glutaraldehyde/0.1M cacodylate. Based on my early readings, I get the impression that IHC is very difficult/impossible with glutaraldehyde fixed tissue, as the fixative denatures antigens irreversibly.

Does anybody have any experience with this, and if so could you offer me some guidance? Thanks.

Many fixatives denature antigens irreversibly - mostly through cross-linking. Have you tried any antigen retrieval methods?

Thanks for your reply. I haven't tried anything yet. I'm still in the exploratory phase, but I knew some retrieval method would be necessary. Is there a method/protocol you'd recommend?

No methods in particular off the top of my head. However, basically they boil down to three types - heat (usually heating to a defined temperature in a buffer), chemical (basically partial chemical digestion of the proteins), and enzymatic (protease based, works by partial digestion of the proteins). Which one you should use, depends on the tissue you are using, what you want to see, and the antibody you are using.

I have had good success with protease digestion in the past, but it can be very finicky to optimise as precise timing is required.