Does anyone have a protocol for tubulin staining in cells transfected with a
GFP-fusion protein.  We see a cytoplasmic localization of the GFP fusion
protein, and it looks filamentous similar to tubulin.  We would like to stain
for tubulin with a red fluorescent probe so that we could check for
co-localization.  I'm worried about losing our GFP signal during the fixation
needed for tubulin staining.  Most of the protocols that I have seen using
monoclonal anti b-tubulin require fixing in methanol, ethanol or acetone which
destroys the GFP.  We tried the red-orange fluorescent BODIPY 564/570 paclitaxel
from Molecular Probes, but we didn't routinely get a filamentous pattern.  Any
help and/or suggested protocols would be greatly appreciated.

Susan Garfield
Laboratory of Experimental Carcinogenesis
Division of Basic Sciences
National Cancer Institute, NIH
Building 37, Room 3C28
37 Convent Drive MSC4255
Bethesda, MD  20892-4255
Phone: 301-496-5688; Fax: 301-496-0734