Acid Native PAGE

Jeremy Murray jez at
Sat Nov 18 20:41:51 EST 2000


kim et al. (2000) Analytical Biochem. 282: 147

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
> ---

Jeremy M. Murray               E-mail: jez at
Dept. Molecular Biophysics & Biochemistry
Yale University
325 Bass Center
PO Box 208114
New Haven
CT 06520-8114                  ph/fax:(203) 432-5280/3104    

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