Post subject: Polyethylene glycol embedded human tissue
I have been using immunocytochemistry for year. Recently, I received human sections (prefrontal cortex) embedded with polyethylene glycol from a brain bank. I tried all tricks that I know to label fibers with antibodies against tryptophan hydroxylase. Unfortunately, I didn’t get any staining. I have used the same antibodies before in formalin-fixed tissue and paraffin-embedded tissue sections from human, rat and mouse without any problem. I will appreciate any suggestions.
Mon Mar 19, 2012 7:59 pm
Joined: Oct 11, 2004
Post subject: Hi Jimmy
I assume that you cut the sections?
You used the PEG protocols?
( No Ag retrieval reqd).
Also, to get an assurance that your PEG immunomethod is just as good as your usual methods, you immunostained with an irrelevant primary Ab, like anti GFAP?
I never liked PEG and thought it a tedious waste of time....sure, the morphology is very nice.
I would re-embed in Pwax, if it were my tissue block.
Tue Mar 20, 2012 7:10 am
Joined: Jan 18, 2011
Thanks for your feedback. No, I did not cut the sections. I received them from a brain bank. What I did before running immunohistochemistry was to rehydrate the sections in 95%, 75% and 50% ethanol. To remove the embedding matrix, I incubated the sections 30 minutes in a water bath (62°C) in 50% ethanol. After that I processed for immunohistochemistry. I tried antigen retrieval but I did not see any difference. I tried an anti-GFAP and got nice astrocytes.
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