[Histonet] Axl IHC for frozen sections

From:Christian Franci

I got several questions and would much appreciate any help you can give me.

1) Heard that pickling tissues over night (in 30% sucrose at 4 degrees C)
prior to freezing them works well to limit the size of ice crystals forming
in the specimens.  MY concern is over degradation of my tissue  during this
time.  Since, I'm working with precious tumor xenographs, should I be
concerned that this might happen?  Any way to prevent this?  Also, after the
sucrose bath, is there a difference as to how one freezes the samples?

2)Lastly, I need to find antibodies specific for staining MOUSE fibroblasts
as well as for HUMAN Axl (aka: UFO,ARK)
Anyone have any experience with either and, can recommend a good supplier?
The less species cross-reactivity the better but, I'd settle for less non
specific staining. Would prefer monoclonals  but, if there are good
polyclonals I'll gladly try them out so long as I can get an idea as to how
much to use to stain my sections.

Thank you all for all your help with the CD31 staining problems I was
having.  Got some great replies and sage advice and I'm happy to report that
it all work out well.

Thanks again,
Cheers
Chris


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