I need to detect p53 in murine E15.5 embryos that overexpress this protein. The embryos were collected and fixed overnight in 10% Neutral Buffered Formalin. Then, I cryopreserved these embryos in 30% sucrose until they sunk to the bottom of the tube and embedded them in OCT. I took sections around 14um in positive charge slides. These slides were stored at -80C. I'm trying to use these slides for p53 IHC using CM5 antibody but no p53 has been detected. Does anyone has protocol for p53 IHC in frozen sections? Thank you in advance!
See attached reprint and the standard protocol for frozen section IHC for my lab. Hopefully these help.
Dave Sherry
Dear Joselyn:
Your sections were not frozen sections because your embryos were collected and fixed overnight in 10% Neutral Buffered Formalin before samples were embedded in OCT.
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