Hello. I am a novice in immunohistochemistry. I have been trying to
stain some rat hearts with PCNA (PC10). I collected the tissue, embedded
it in O.C.T. and stored it at -80C. I use Lymph node as a positive
control. Upon cutting sections, they were fixed in Acetone, then
endogenous peroxidase was blocked with 1% H2O2 in methanol. From then
on, a kit was followed. After getting no results, I began calling around
and someone mentioned that this antibody could not be used for frozen
sections due to antigen solubility. Well, I obtained some paraffin
embedded human tonsil and tried the procedure and it worked fine. Could
someone please enlightened me on antigen solubility? What is it soluble
in? Is there something I can modify so that I can use my frozen tissue?
Thanks in advance
-chris
