Silver Staining and BME

Achim Recktenwald achim at ibex.ca
Fri Sep 13 13:38:08 EST 1996


zoltan at bcmp.med.harvard.edu wrote:
> 
> I'm looking for a good solution to an old problem - those nasty
> "pseudo-bands" in the 70kDa range which can plague silver-stained
> protein gels.  I recently finished off a bottle of beta-mercaptoethanol
> which I had been using to prepare loading buffers for SDS-PAGE with
> excellent results, and since I have started using other batches
> (including a brand new bottle of BioRad electrophoresis grade) I have
> had excessive staining around 70 kDa in all gel lanes.  In addition to
> trying various brands and batches of BME I have also tried lowering the
> concentration of BME in the samples, but I've had no luck.  Can anyone
> help me out here?  I would appreciate replies by email as well as to the
> newsgroup.  Thanks!
>                        - Eric Arn
>                        zoltan at bcmp.med.harvard.edu


Your sample-buffer may be too old:  make fresh one and leave the
bromphenolblue out of it;
I usually make a larger stock and freeze it in Eppendorf tubes. The
marker-stain is only added at the last moment. I only use this
SDS-buffer-bromphenol-mixture for less than a week.

Achim



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