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
Full Version: DAPI/Hoechst vital staining
QUOTE (neopeg @ Nov 2 2009, 07:31 AM) 
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?
Fixing shouldn't kill the GFP, it is pretty stable... have you tried 2% cold paraformaldehyde?
DRAQ 5
http://www.biostatus.com/product/draq5/
however, I agree with above commentary. What are your fixation conditions?
http://www.biostatus.com/product/draq5/
however, I agree with above commentary. What are your fixation conditions?
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