SDS-PAGE running problem, help!

Dr. Artem Evdokimov eudokima at mail.ncifcrf.gov
Wed Jul 18 12:04:10 EST 2001


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  |
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