Using BrdU as a DNA-replication label requires to denature the DNA before the
detection with Antibodies to make the antigen accessible. Two methods are
commonly used: Denaturation with HCl and denaturation with NaOH. Unfortunately,
both methods will kill a GFP signal in the cells due to the pH-shift.

Has anybody successfully detected BrdU- without destroying a GFP-signal in the
cells? I am thinking about trying a denaturation with formamide and heat,
simillar to what is used for FISH, but I wonder whether anybody has tried that
already with or without success.

Steffen


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Dr. Steffen Dietzel
Dept. of Cell and Structural Biology
University of Illinois at Urbana-Champaign
601 S. Goodwin Ave.
Urbana, IL 61801
Phone: +1/217/333-8372  Fax: +1/217/244-1648
e-mail: [log in to unmask]  Web:  http://www.life.uiuc.edu/belmont/dietzel
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