HELP: extracting and digesting 2D-GEL spot
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.
Dr. Simone Cristoni
University of Milano - (CISI)
Via Fratelli Cervi 93
20090 Segrate, Milano (Italy)
E-MAIL: dtoycr at tin.it
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