Irregular bands with my protein gels

Andy Phillips andy.phillips at bbsrc.ac.uk
Wed May 7 05:59:50 EST 1997


packard at kersur.net 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
> 
> 

I vaguely remember an article by Ed Southern in an old Methods in
Enzymology (about 1981?) concerning vertical agarose gels for DNA
separation. He mentioned that the glycerol usually added to loading dye
makes the DNA samples run as dumbells, as it makes the sample stick to
the walls of the well. His cure to this was to use an alternative to the
glycerol - I seem to remember (this was 17 years ago!) that he used 10%
Ficoll. Whether this would work in an SDS gel is another matter - is
Ficoll heat stable?

I tried, but failed, to find this article in my reprint collection, and
ISI doesn't go back far enough.

Andy

Andy



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