HELP: extracting and digesting 2D-GEL spot

Simon dtoycr at tin.it
Thu Sep 19 12:02:41 EST 2002


I am an Italian biological researcher working on protein and peptide
molecules by mass spectrometry. Recently I am trying to analyze some protein
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 to
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 in
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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