GFP and BrdU staining

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


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



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