Acid Native PAGE

Michael Witty mw132 at mole.bio.cam.ac.uk
Wed Nov 8 05:16:09 EST 2000


I have never tried it, but could adding a little glutaraldehyde make the
complex stick together and be easier to handle?  Mike.

On 7 Nov 2000, lou wrote:

> I have two proteins, one with a pI of 8.8 (Mw=50kDa) and the other with
> a pI of 8.3 (Mw=90kDa) I am trying to make a complex of the two proteins
> (which should be irreversible) and would like to see this complex
> formation, preferably on a native gel.
> 
> I have attempted to run an acid native PAGE,. I have made my gels (25mM
> Tris Glycine, 12% Acrylamide) and the buffer is Tris (25mM)  Alanine,
> pH7.4. The buffer that the samples are in vary but all are 25mM buffer
> with 100mM NaCl. The loading dye contains glycerol and methyl green.
> 
> The problem that I am having is that the samples do not run very far and
> the bands are quite broad. I have heard that lowering the salt
> concentration of the samples can help with resolution, but by how much.
> Also does anyone know if the pH that I have tried is low enough?? Should
> I just try another gel at lower pH?? Is there any other way in which
> people might be able to advise me??
> 
> I hope that someone can help
> Thanks
> 
> Lou
> 
> 
> 
> ---
> 
> 






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