problem with PAGE_SDS

didier d-fesque at nimr.mrc.ac.uk
Wed May 7 05:54:28 EST 1997


you wrote

>I'm having some problems with irregular bands on my protein gels. I'm
running a purified antigen (approx. 60 KD) using a 10% acrylamide gel.
The problem is that after staining/destaining my bands appear dumbell
shaped. This doesn't happen all the time (about 50 % of the time). I
must adhere to GMP documents..so there are few changes I can make in
the procedure. I was wondering if this is a technique issue or
something real simple that I'm missing. Any help would be greatly
appreciated! Please feel free to email me if you would like more info
about my dilema. Thanks you for your time. My email address is
packard at kersur.net

Kevin M Packard


hi, have a look to the pH of your stacking buffer, the pH of the Tris
solution is instable at 6.8, this induce change in the migration of
protein during PAGE. the simple way to avoid this is to freeze aliquot
of thius buffer (20ml) and after using this aliquot store it at 4 degre
, for a few week!

didier



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