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
0
3 replies to this topic
#2
gfischer
-
- Global Moderators
-
- 179 posts
Veteran
7
Neutral
Neutral
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
Above all things, if kindness is your king,
then heaven will be yours, before you meet your end
then heaven will be yours, before you meet your end
#3
bob1
-
- Global Moderators
-
- 4,337 posts
Thelymitra pulchella
222
Excellent
Excellent
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
-
- Active Members
-
- 19 posts
member
0
Neutral
Neutral
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?
