SilverQuest Troubleshooting

[Dr Engelbert Buxbaum]

Am 20.09.2007, 11:10 Uhr, schrieb Arne Christensen <arnec from bio.umass.edu>:

> I'm using the SilverQuest Silver Stain kit and I'm reproducibly having a  
> curious problem.
>
> I run the staining protocol one time, and get a negative stain of  
> protein bands.  Protein is not overloaded, this will be made clear  
> later.  The two bands that do stain however, are the 75 and 25 kDa  
> prestained (only two bands stained pink) molecular weight markers.
>
> Because the staining does not work, I destain with sodium  
> thiosulfate/ferranocyanide.
>
> When I follow the same protocol a second time, the staining works  
> great.  No negative stain, suggesting that the protein was not  
> overloaded.  All markers stain well.  This has been the case 4 times.
>
> Has anybody had a similar problem, or is familiar enough with the  
> chemistry of silver staining to recognize the problem?

This has been reported in the literature before, I do not recall a  
specific reference however. Silver staining is an iffy procedure with all  
kinds of strange results. Fluorescent staining is easier and more  
reproducible. Staining kits are available for example from Invitrogene  
(formerly Molecular Probes), but check also

@article{Rab-01,
         AUTHOR= {T. Rabilloud and J.-M. Strub and S. Luche and A. van  
Dorsselaer and J. Lunardi},
         TITLE= {A comparison between Sypro Ruby and ruthenium(II) tris  
(bathophenanthroline disulfonate) as fluorescent stains for protein  
detection in gels},
         JOURNAL= {Proteomics},
         VOLUME= {1},
         YEAR= {2001},
         PAGES= {699-704},
         LANGUAGE= {engl}
}