I have some adrenal tissues that I want to check the localization and expression levels of some receptors. I have lost the paraffin-embedded blocks, but I have the formalin-fixed tissues in containers. I do not know whether antigen retrieval would recover most of the proteins or it would be a total waste of time and resources. Kindly advise.
Probably you do not have an easy way to obtain another new tissue. So, I recommend you process the pieces that you have in buffered formalin, and make a HE stain to see how the morphology was. For some stable proteins such as the cytoskeleton and extracellular matrix, you will probably get good results for immunofluorescence. Lectins are also a good option. Depends on what you would like to study further. If you desire some nuclear stains or phosphoproteins, or to detect RNA or DNA such as FISH, I really recommend starting from the beginning.
Thank you, Paula. I appreciate your advice. To be specific, I want to assay TNFR1 and MC2R using primary and conjugated secondary antibodies and laser scanning confocal microscope.
You can also try antigene retievel in the microwave especially when you are working with paffin sections. And you can use more sensitive detection systems like biotinylated secondary antibodies and Streptavidin (3 step method)
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