is GFP compatible with BrdU immunofluorescence

Alfredo Pagliuca pagliuca at cds.unina.it
Fri Dec 18 10:46:11 EST 1998


Hi everybody,
i've just encountered a problem with GFP:I use a GFP expression vector as
a marker for transfected cells, but as I am performing immunofluorescence
experiments, I have also noticed that when my sample (cells growing on
coverslips) is fixed with ethanol/methanol at -20°C no GFP is detectable.
I was  told that fixing the samples with 4% paraforamdehyde might work
fine, however I fear this might not be compatible with  the following
assay for BrdU incorporation, in which the cells must be subjected to pH=2
for DNA denaturation. So basically the question is: are there any methods
for BrdU assay which allow an alternative fixation method of the samples?
Are there an tricks for preserving GFP activity in the fixation
conditions? If anybody knows, I would be very pleased to hear from, as it
seems I ended in a cul-de-sac.
Bye
Alfredo Pagliuca
PhD student
University of Naples - Italy



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