Silver stained gels - background problem when using DTT

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Fri Jan 9 11:44:59 EST 2004


Amanda wrote:

> I am having a small issue with my silver stained gels. (Mini gels, 1.5mm,
> run at 18mA, unlimiting voltage).
> 
> I use a reducing treatment buffer containing DTT as our supplies of
> b-mercaptoethanol are badly contaminated with keratins. However, I get a
> horrible uniform background yellow/brown colour (during the development) in
> the lanes I loaded with this buffer (I've tried just buffer alone).


Which staining protocoll are you using? Have you tried Heukeshoven &
Dernik? In my hands that gives the best results from all the methods I
tried. 

Also make sure that all your reagents are scrupoulously clean.

You may consider replacing silver staining with fluorescent staining
(nile red or one of Molecular Probes Sypro stains). They are almost as
sensitive as silver.



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