IUBio

Good silver staining kit

Artem Evdokimov AEVDOKIMOZ at cinci.rr.com
Wed May 8 19:08:35 EST 2002


Are you sure that a 'homebrew' silver stain isn't good enough for what
you're doing ?

For example, here's an approximate protocol that we have developed cause we
hate waiting for silver stain. It's quick-n-dirty (that is it gives slightly
higher background and slightly less sensitivity than full-blown 6-hour
protocol) but it works well enough for most of our purposes.

1) fix gel (microwave in your favorite AcOH/MeOH/H2O mixture for 1 min on
high)
2) Wash gel (microwave 1 minute 3 times in three changes of H2O)
3) <1 min microwave in 0.1% AgNO3 (heat to at least 70C, but do NOT let it
boil!)
4) Wash gel (microwave 1 minute 2 times in two changes of H2O)
5) <1 min microwave in 0.2% Na2S2O3 (heat, but do NOT let it boil!)
6) Wash gel (microwave 1 minute 2 times in two changes of H2O)
7) Develop gel with standard carbonate-HCHO mixture (look at bands
appearing, stop when ready with excess of AcOH/MeOH/H2O mixture). Generally
under 2 minutes.
8) store in AcOH/MeOH/H2O mixture or if needed, gently destain with acidic
thiosulphate.

Total time needed - well under 30 minutes.

A.G.E.

"Froog" <froogly at hotmail.com> wrote in message
news:f30508df.0205080856.7dd31056 at posting.google.com...
> Hi all
>
> I am new to proteinland and I would like to know if one of the silver
> staining kits on the market is used more than others (ie like Qiagen
> for plasmid preps for example) - or if people can recommend a reliable
> kit.
>
> Thanks very much in advance
>
> Claire
> Sheffield, UK





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