silver stain problem

Artem Evdokimov AEVDOKIMOZ at cinci.rr.com
Sat Oct 12 22:06:06 EST 2002


Glad to hear that it worked for you :) If you like to experiment with fun
stuff, there's a relatively novel approach to protein gel staining that
involves incubation of the gel in the presence of TCA or chloroform,
followed by wash and UV irradiation (just your regular UV transilluminator).
After 20-30 minutes of UV irradiation, protein-containing bands will become
fluorescent (greenish-yellow, visible in the dark) due to photoreaction of
the chloro-organics with Trp residues. This method is about as sensitive as
Coomassie, and if you have no Trp then it won't work, but it's something fun
to try if you have a slow day in the lab. There is a nice recent reference
describing the technique but I can't quite remember what journal etc. was it
in, sorry.

Artem

"janosch groening" <janosch.groening at ioez.tu-freiberg.de> wrote in message
news:3DA6F9A8.E55CE15F at ioez.tu-freiberg.de...
> Hello A.G.E.,
> as coincidence works, I did just yesterday for the first time the
"Quick&dirty"
> silver staining method (microwave) which you mentioned here a few months
back.
> I had two SDS-PAGE's, one stained with "my" usual method (3h) and the
other one
> with the quick method (30min). It worked really great, at the first
approach.
> And there were not such _big_ differences between the quick and the "slow"
> method.
> Actually, though, I did first the step with the thiosulfate and then the
> staining-step with the silver, as "my" "old" method is proceeding like
that.
> just wanted to take the advantage to "report" this, seeing your post today
;).
> with kindest regards
> janosch
>
> Artem Evdokimov schrieb:
>
> > Most likely poor wash in between changes of reagents, or you have
> > inadvertently heated the gel at the developing stage.
> >
> > A.G.E.
> > <beismann at em.uni-frankfurt.de> wrote in message
> > news:20021010142432.2660.qmail at ww02.hostica.com...
> > > Hello,
> > >
> > > I have done silver stains of SDS gels many times before but last time
> > something strange occured: I got an inverse stain with dark background
and
> > light protein bands (altogether rather faint). Before this stain, I
prepared
> > fresh thiosulfate buffer, and formaldehyde was freshly added to the
silver
> > solution and the developing solution.
> > > What can be the cause of the inverse staining, an thus, how can I get
back
> > positive stains?
> > >
> > > Thanks for helping me,
> > >
> > > Silke
> > >
> > >
> > >
> > >
> >
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>





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