[Histonet] Unable to detect protein tagged to GFP

Samuel Beckerman 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 
and mouse"

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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