In article <4rtpch$5at at mserv1.dl.ac.uk>, Peter Corish
<pcor at bioch.ox.ac.uk> wrote:
> Has anybody had any experience (good or bad) using the advertised
> Clontech polyclonal GFP antibody for immunoprecipitations or Western
> blots? In particular, how specific has it been and what level of
> background has been observed? We are interested in using it to look at
> GFP half-life, so any comments would be greatly appreciated.
I've used it for Westerns and immunofluorescence. Works fine in Westerns,
but the batch I got was a bit dirty--I had to clean it up by pre-adsorbing
with acetone powder. Once I did that, it give crisp and clear detection
of a single band (presumably GFP !). I could get away with a 1:10,000
dilution for a Western dectecting GFP expressed in E.coli.
In immunofluorescence in mammalian cells, the background seems quite low. See
http://www.embl-heidelberg.de/~jwhite/project/prelim/preliminary.html
for a couple of immunofluorescent images with the anti-GFP antibody.
HTH,
jamie
--
Jamie White
Light Microscopy Group
Cell Biophysics Programme
European Molecular Biology Laboratory
finger jwhite at phenix.embl-heidelberg.de
