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Hi all,

not strictly confocal question

We´re using antibodies to determine CREB phosphorylation and translocation
of NFAT to nuclei of glandular (exocrine pancreas) and smooth muscle
cells. After several weeks changing Ab dilution, incubation time, block
solution,...we can´t see in the nuclei a decent signal of phosphorylated
CREB (supposed the only place for this molecule) or NFAT. We suspect that
we are simply failing to detect the presence of labelled NFAT/CREB in the
nuclei (NFAT in cytosol decreases when we apply stimuli known to induce
nuclear accumulation)
Is it possible that some cell types have a more "impermeant" nuclear
membrane? We have already prolongued the incubation time in triton.
Any idea would be wellcome

Thanks


--
Dr Pedro J Camello
Dpt Physiology
Faculty of Veterinary Sciences
University of Extremadura
10071 Caceres
Spain
Ph: 927257100 Extension 1321
Fax:927257110