PAGE gels- can you make up to 20% with Tris/Glycine?

Kresten kresten at my-deja.com
Wed Feb 9 12:20:44 EST 2000


No problems w. 20% gels. They can be a bit more difficult to dry
(cracking), so if you really need to do this you could soak them in some
glycerol before drying them.

I am working with a 10 kDa protein. When I run that on a 20%T 2.6% C
SDS-PA gel the protein band is approximately in the middle of the
running gel when the tracking dye runs out. So with a 17 kDa protein
you will probably have to run the gels some more after the dye runs out
in order to get max separation around 17 kDa.

Kresten

In article <38A05A02.C3BBD530 at arches.uga.edu>,
  Keith Pitts <k20man at arches.uga.edu> wrote:
> We generally use a standard homemade 12% Tris/Glycine SDS-PAGE gel
> procedure (from Current Protocols) for our proteins, but are trying to
> get tighter separation.  I have seen recipes for up to 15% acrylamide
> gels in the glycine system.  Does anyone know if you can go up to 20%
> or
> so in this system?  What, if any consequences are there, and how does
> it
> affect the separation of the proteins?  We are looking at a ~17kD
> DNA-binding protein and would like to save tricine buffer recipes/gels
> as a last resort as they would be significantly less convenient.
>
> Thanks much
> Keith
>
> --
> :-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:-:
> Keith Pitts
> Research Assistant            Univ. of Georgia
> k20man at arches.uga.edu
>

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Kresten Lindorff Larsen, Dept. Yeast Genetics


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