SDS-PAGE running problem, help!
mw132 at mole.bio.cam.ac.uk
Wed Jul 18 12:19:40 EST 2001
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.
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 ?
> 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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