SDS-PAGE running problem, help!

Michael Witty mw132 at mole.bio.cam.ac.uk
Wed Jul 18 12:19:40 EST 2001


Dear Amanda,
           is something growing in the bottle you store electrode buffer
in?  We had a similar problem in our lab once and solved it by cleaning
the bottle (which we continually recycled without washing) very well.

Mike.

On Wed, 18 Jul 2001, Dr. Artem Evdokimov wrote:

> Could you describe you rgel in more detail ? Is it a pre-cast gel  or
> did you pour it yourself ? What % AA ? What voltage/current ? For how
> long ? What are you loading ? What are the buffers ?
>
> A.
>
> Amanda Cage wrote:
> >
> > I have problems running SDS-PAGE gels because a pencil-like line runs
> > continuously from one end of the gel to the other end of the gel
> > horizontally at about 3/4 of the way down on the gel when I do the
> > silver staining.  I am not sure why that happens and do not know how
> > to get rid of it.  The strange thing is when I use commassie blue to
> > stain a different gel (not the same gel as silver but contain the same
> > samples), a color gradient appear across the gel.  The top appear
> > almost colorless.  The middle of the gel appears a light blue.  The
> > bottom part of the gel (3/4 way down) appears a deeper blue than the
> > middle.  I could not get the entire gel to stain a uniform blue color.
> >  Could the problems with these two gels be the running buffer
> > contaminants or is it something else?  I washed the SDS off the gel
> > very well so I believe that is not the problem.  Please help!  Thank
> > you.
>
> --
> |Dr. Artem Evdokimov   Protein Engineering |
> | NCI-Frederick        Tel. (301)846-5401  |
> |              FAX (301)846-7148           |
> |        eudokima at mail.ncifcrf.gov         |
> |      http://mcl1.ncifcrf.gov/plague      |
>




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