[Histonet] Unable to detect protein tagged to GFP
sbecke2 <@t> emory.edu
Mon Jun 15 16:27:02 CDT 2009
This is a follow up to posts made in February of this year under the
heading "[Histonet] expression pattern of GFP fusion protein in cells
My name is Sam Beckerman. I am currently conducting research at Emory
University. Recently, my group has experienced difficulties detecting a
GFP-tagged fusion protein in neuronal cells. Specifically, we have made
lentivirus that expresses GFP tagged to our protein of interest which
we inject into rat CNS.
After fixation (4% PFA) and cryosectioning we have preformed
preliminary examination for GFP and are able to detect strong GFP
signal in our tissue. However, we have been unsuccessful at using
antibodies against our protein to co-localize with GFP.
We have used this same virus to infect 293 cells and PC12 cells and
have been able to colocalize our proteins in vitro.
Any help would be much appreciated.
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