Immunoprecipitation

Jamie White jwhite at embl-heidelberg.de
Sun Jul 14 08:45:00 EST 1996


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



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