immunofluorescence of parafin embedded tumor section - (Dec/17/2008 )
Could some one in the forum please point me to the link on the forum or share there protocol for double imunofluorescence of parafin embedded human tissue sections please. Especially points to take care.
I'm not sure if there is an actual protocol, I'm sure you can find it online though. Just Google Immunofluorescence and parafin.
In my experience the immunofluorescence was much much better on cryosections, so that's what we do in our lab. Parafin embedded tissues are great to keep the structure, but the parafin also covers/changes some cell surface markes, which is what I'm looking at. If you do frozen sections with the cryostat I didn't have that problem.
You'll also have to be careful to remove all the excess aldehydes from the sections as these will autofluoresce and interfere with the antibody mediated reactions.