[Histonet] IHC on 3 year fixed tissue?
kevinpe <@t> mail.med.upenn.edu
Sat Mar 13 13:15:12 CST 2010
I've been asked to perform anti-GFP IHC on some mouse bones which have
been sitting in NBF for 3-4 years. My Abcam rabbit polyclonal gets great
results in GFP mouse control tissue, but I can't seem to get any results
out of these bones. I've tried citrate buffer HIER using autoclave,
hotplate, and 65 degree water batch over night, but my signal is
non-existent (or really the noise is so huge you can't distinguish).Do
the great minds of histonet have any ideas on how to achieve good
staining? Is enzymatic epitope retrieval worth a try? Does anyone have
a time machine I can use to stop my predecessors from just sticking the
bones in NBF and forgetting about them for years?
This listserv has helped me countless times, I am truly appreciative of
the knowledge and experience available on Histonet.
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