western blotting (PVDF) of EGF receptor

David A Rintoul drintoul at matt.ksu.ksu.edu
Mon Oct 14 14:53:12 EST 1991


  If anyone out there has a tried and true recipe for
transfer of SDS-PAGEd proteins containing the EGF
receptor (from A431 in our case) to a PVDF membrane
for western blotting, I would be happy to hear from
you.  I am not a protein or nucleic acid chemist,
but our lab is interested in the effects of the
lipid environment on the activity of the EGF receptor
(see Song, et al, JBC (1991) 266:10174-81), and
thus have to become quasi-adept in protein alchemy
such as western blots.  This seems to be an area where
somewhat arbitrary changes in buffers, times, voltages,
pre-soaking of the gel or membrane, etc can make the
difference between success or failure.  Thus far we
have failed.  So, if you know of a recipe, or a reference
for a recipe that will help us transfer the EGF receptor
to PVDF, I'd love to hear from you.  Thanks in advance.
Please e-mail to me: DRINTOUL at matt.ksu.ksu.edu.
 

-- 
Dave Rintoul            Internet: drintoul at matt.ksu.ksu.edu
Biology Division - KSU       BITNET: drintoul at ksuvm.ksu.edu
Manhattan KS 66506-4901                         TCN: id2418
(913)-532-5832 or 6663                  FAX: (913)-532-6653



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