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DAPI/Hoechst vital staining


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#1 neopeg

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Posted 02 November 2009 - 05:31 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

#2 gfischer

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Posted 02 November 2009 - 02:08 PM

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

It's an extra few steps, but what about using an antibody for GFP?
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#3 bob1

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Posted 02 November 2009 - 04:40 PM

Fixing shouldn't kill the GFP, it is pretty stable... have you tried 2% cold paraformaldehyde?

#4 blotted

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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?




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