Dick,
We have been using the method since the paper came out with great success.
Prior to this destaining, our success rate with silver stained samples
(including M. Mann's silver staining protocol) was poor. Using 1D gels and BSA
loading studies, we were at least ~5-10 times less sensitive with silver stained
samples as compared to coomassie. Once we started using Farzin's destaining, we
are at least as senisitive with the silver as coomassie. The method is easy and
very fast (typically, the gel band turns opaque within a few seconds of contact
with the thiosulfate/ferricyanide solution). The only danger is that it is hard
to see the gel piece once it is destained so good eyes are a requirement to
avoid sucking it out with the wash solution.
Dale
Dick Wicks <dick@fortron.com> on 08/19/99 09:21:55 AM
To: Recipients of ABRF List <abrf@aecom.yu.edu>
cc: (bcc: Dale Patterson/HQProdDev/PBIO)
Subject: Silver destaining method for MALDI analysis
Hello ABRFers:
Has anyone looked at the ferricyanide/thiosulfate method to
destain silver stained proteins from 2D gels before MALDI
analysis? (Gharahdaghi et. al. Electrophoresis 20: 601, 1999). I
would be especially interested in any side by side (destain vs. no
destain) studies. Thanks.
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