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GFP cells intracellular staining


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#1 MNR

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Posted 11 April 2011 - 11:07 AM

I am kind of desperate because I would like to look at cytokines production by GFP cells. But when I am doing the fixation and permeabilization protocol from eBioscience I am loosing all my GFP staining.
What are you advices ? How am I suppose to fix and permeabilize my cells for that ?
Thank you in advance

#2 Rsm

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Posted 11 April 2011 - 10:25 PM

Fixation almost always destroys GFP signal. How about you sort your (live) cells for GFP, and then do a fix and stain on the isolated populations? You could also try an anti-GFP antibody conjugated with some dye, for example FITC conjugated. That should work on fixed cells.
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