I want to isolate and analyze mRNA for different targets from only the part of tissue which stains positive for a specific protein. I know that RNA can be isolated from snap frozen tissue, but since it degrades so easily, is it possible to collect any after immunohistochemical staining? I need the mRNA to do microarray analysis/westerns for expression of multiple targets so that I can avoid optimizing the antigen retrieval methods for the multiple targets i need to study.
I know that you can perform Laser Capture Microdisection (http://en.wikipedia.org/wiki/Laser_capture_microdissection) to Isolate the cells of interest from a stained IHC section. I understand it is pretty tricky though and even though frozen or paraffin embedded tissue could be used there are pros and cons to both. RNA will degrade easily but then fixation can effect the quality of RNA too.
Thanks..didn't know such a method existed...!
but i think i got it .. i could isolate the different cells using different cell markers (magnetic sorting) and isolate RNA then instead of doing an IHC..
Yes, that would definitely be much easier. You could even FACS sort if possible. Laser Capture is most often used when the tissue is already sectioned...
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