I want to resolve and separate p67 and p64 c-Myc protein by SDS-PAGE to recapitulate the below shown figure from the below linked paper. In the paper they used 10% polyacrylamide to separate p67 and p64 c-Myc, but the separation wasn't very good. Is there a more optimal concentration that may work better? Alternately, would a gradient gel (4-15%, 4-20%, 8-16%, or Biorad AnyKD) work better?
http://www.ncbi.nlm.nih.gov/pubmed/26177060
Gradient gels wil definitely improve the resolution, however, I think the Tris-Tricine system might be a good choice for your purpose. http://www.nature.com/nprot/journal/v1/n1/full/nprot.2006.4.html
It provides really great separation (and not only at extremes of Mw) with nice-looking narrow bands. The only critical thing - no leakage between upper and lower buffers, otherwise the resolution will be dramatically compromised.
Hi,
I would like to draw your attention that the resolution of the gel you've showed is pretty good. getting a separation of 5% of the MW of the protein in gel electrophoresis is as good as it comes.
Cheers,
David.
Dear Brian,
Although the resolution looks good in your case as David said,but even if you want to resolve more, then use the gradient of 4-15 %,and use LDS rather than SDS and try to do the electrophoresis at 4 degree C.
All the best.
- Badges
- Science method