How to fix GFP expressing cell to preserve GFP signal in immunofluorescent stain - (Apr/01/2009 )
Hi,
I am trying to do a Brdu staining on Spinal cord sections that have GFP cells, the purpose is to see if those cells have been proliferating because of a disease or not. since I have very few good sections( they are PFA perfused, frozen sections), I am afraid I might loose GFP because of harsh treatment that involves Brdu staining, does any one has any suggestion?
Bneuroscience on Apr 1 2009, 04:37 PM said:
I am trying to do a Brdu staining on Spinal cord sections that have GFP cells, the purpose is to see if those cells have been proliferating because of a disease or not. since I have very few good sections( they are PFA perfused, frozen sections), I am afraid I might loose GFP because of harsh treatment that involves Brdu staining, does any one has any suggestion?
Not sure what harsh treament BrdU staining involves, but in worst case scenario, couldnt you take a picture of the GFP and overlay with the picture you take of the BrdU after the staining procedure ?
Thanks for taking time and reading my post/The acid treatment, kills the GFP. The journal wants to see the co-localization on the same picture.
Bneuroscience:
The acid treatment is presumably killing the fluorescence, but not the GFP itself. That being the case, you can use an anti-GFP antibody (of which there are plenty). The antibody should still be able to bind, then you can use a green secondary to reproduce it's natural green-ness!
-Carlton