GFP and BrdU staining

Mayumi Naramura (MNARAMURA@atlas.niaid.nih.gov)
Thu, 20 Feb 1997 14:33:00 -0500

Hi,
Does anyone have experience staining GFP (S65T, humanized codon, not a
fusion protein)-expressing cells for BrdU incorporation? I'm afraid
standard alcohol fixation and HCl treatment is too harsh for GFP. I
read about nuclease treatment and UV breaking. Which anti-BrdU
antibody would you recommend?
In fact, is there a better way to detect proliferating cells? Is
there, say, a thymidine analog that is incorporated into newly
synthesized DNA and fluoresces on its own? Certain easily-detectable
antigen that is "really" specific to some cell cycle stage?
Mayumi Naramura, M.D.
NIH/NIAID
Lab of Immunology
Twinbrook II, Rm. 125
12441 Parklawn Drive
Rockville, MD 20852
Phone: 301-402-4595
FAX: 301-594-2522
e-mail:mnaramura@atlas.niaid.nih.gov