extracting and digesting 2D-GEL spot

Pier Carlo Montecucchi pcmontecucchi at compuserve.com
Sun Sep 22 11:54:44 EST 2002


Sometimes, I'm using the isoelectrofocusing technology for protein / peptide
isolation with the M.I.E.R. apparatus developed by the Italian company
Talent (Trieste).

The system works.

On the isolated components, MS or Edman degradation.


Pier Carlo

Pier Carlo Montecucchi, C.P.C., C.C.
Via delle Rosine n. 1
10123 Torino
Email: pcmontecucchi at compuserve.com

"Simon" <dtoycr at tin.it> wrote in message
news:Rqni9.115054$ub2.2449565 at news1.tin.it...
> I am an Italian biological researcher working on protein and peptide
> molecules by mass spectrometry. Recently I am trying to analyze some
> extracted from GEL spot. I have some problem to solve:
> 1) Do you know if exist a protocol for extracting the protein from the GEL
> spot without digest them?
> 2) Does this protocol work for any kind of protein?
> 3) I am trying to digest the protein directly in GEL. Many protocols say
> dry the gel and reydrate with trypsin solution. My question is: it is
> possible that the trypsin does not go inside the GEL but remains outside
> solution? I have followed various protocols but at the end seems that the
> peptides do not exit from the gel. For this reason I have thought that the
> digestion does not take place.
> I would like to thank you very much for your help.
> Sincerely yours
> Simone Cristoni
> Dr. Simone Cristoni
> University of Milano - (CISI)
> Via Fratelli Cervi 93
> 20090 Segrate, Milano (Italy)
> Tel: +39-02-50330426
> FAX: +39-02-50330414
> E-MAIL: dtoycr at tin.it

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