I have assessed the specificity of a particular monoclonal antibody against a virus by immunofluorescence. I'd like to further validate it by testing it by IHC. Therefore I'd like to infect cells in culture, and then inject them in a piece of fresh organ, fix in formalin, embed in paraffin and then cut slides before performing IHC. Is there any protocol or references to do so ?
$\begingroup$
$\endgroup$
Add a comment
|
1 Answer
$\begingroup$
$\endgroup$
1
Injecting into a fresh organ seems like a needless complication if you're just validating your antibody and not interested in gathering data about the interaction between the cells and the organ. (In fact, if you're using anything more complex than a flatworm I'd hope your IRB would have problems with needlessly sacrificing animals to get those organs.)
It seems like your purpose would be better fulfilled by making an FFPE cell plug and doing your IHC on that. Here's a protocol for making cell plugs: Johns Hopkins Tissue Microarray Lab Cell Plug Protocol
-
$\begingroup$ Thanks you for your answer. Actually this was a request from a reviewer. Regarding organs I work close to the vet necropsy department therefore getting fresh organs is not an issue. $\endgroup$– CobactanJul 29, 2015 at 21:10