native gel of large complexes

[Virginia Dress]

In article <3kitg3$8c4 at news.ycc.yale.edu>, Danny Chamovitz <chamo> wrote:
> 
> I'm trying to do a native gel of a large protein complex (>550 kDa), but I'm
> having trouble getting it into the gel.  Has anyone tried to use agarose intead
> of acrylamide?  Has this also worked for gel shifts?  Any and all advice will
> be greatly appreciated.
> 
> Danny

A long, long time ago someone in a lab I was working in was using agarose-
acrylamide gels to separate proteoglycans.  I'm not sure if it was
non-denaturing.  So try looking up agarose-acrylamide gels.  You'll need
one
of the glass plates to have a frosted surface (like the writing area on a 
microscope slide) otherwise the gel will slide right out the bottom.  If
you
have a gel rig with an alumina plate (like the mini-gel rigs) that will 
probably work too.  You may be able to pour a vertical gel with agarose if
you have the frosted plates, otherwise I guess you could try it submarine
style.

Ginnie