Hi there;
I wanna stain GFP transfected human cells [adhaerent monolayer culture] with DAPI or Hoechst. My problem is that every fixation kills my GFP in the cells. Does anyone know a good protocol for vital staining of the nucleus without any fixation of the cell.
Thx Neo
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3 replies to this topic
#2
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Posted 02 November 2009 - 02:08 PM
neopeg, on Nov 2 2009, 07:31 AM, said:
Hi there;
I wanna stain GFP transfected human cells [adhaerent monolayer culture] with DAPI or Hoechst. My problem is that every fixation kills my GFP in the cells. Does anyone know a good protocol for vital staining of the nucleus without any fixation of the cell.
Thx Neo
I wanna stain GFP transfected human cells [adhaerent monolayer culture] with DAPI or Hoechst. My problem is that every fixation kills my GFP in the cells. Does anyone know a good protocol for vital staining of the nucleus without any fixation of the cell.
Thx Neo
It's an extra few steps, but what about using an antibody for GFP?
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#3
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Hmmm, I think it's working
Posted 02 November 2009 - 04:40 PM
Fixing shouldn't kill the GFP, it is pretty stable... have you tried 2% cold paraformaldehyde?
#4
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Posted 22 December 2009 - 05:21 AM
DRAQ 5
http://www.biostatus.../product/draq5/
however, I agree with above commentary. What are your fixation conditions?
http://www.biostatus.../product/draq5/
however, I agree with above commentary. What are your fixation conditions?
