From: sandra pilawa <sap@gbf.de>
Date: Sat, 25 Nov 2000 13:03:16 +0100
Subject: silver stain, MALDI-MS, Proteomics
Organization: TU Braunschweig, Germany

Hi,

I am a chemist and work in a proteome research group in germany, but
here-no one could give me an explanation......

I am looking for the reason, why it is difficult to make a maldi MS of
protein spots of a silver stained 2D gel (protein probe, IEF, 2D gel,
silver stain neudorf-methode, cut out, tryptic digest, maldi-ms matrix:
cyano-hydroxy cinnamoic acid). Is the reason the poor amount of protein
(peptide) or is it a problem with the silver ions, ionisation or matrix?

If you have an idea or maybe a paper with a detailed explanation, I am
grateful for all ideas or help.

thanks for your time and ideas
Sandra




******************************************************************************
From: Heino Prinz <heino.prinz@mpi-dortmund.mpg.de>
Date: Mon, 27 Nov 2000 09:48:07 +0100
Subject: Re: silver stain, MALDI-MS, Proteomics
Organization: GWDG, Goettingen

Do you use glutaraldehyd in the protocol? If yes, then this
cross-linker is the most likely explanation. It would interfere
with the action of Trypsin. If no, then you should try to follow
the protocols of the EMBL-group (Matthias Mann, Matthias Wilm, A.
Shevchenko) to the dot. It really works!

Viele Gruesse		Heino Prinz 

Tel.:(++49)-231-133-2480  Fax: (++49)-231-133-1038