Peptide Mass Fingerprinting from PVDF

Marcelo Valle de Sousa (mvsousa@unb.br)
Thu, 15 Jul 1999 17:11:38 +0000

A protein was blotted to PVDF from a SDS-PAGE gel for subsequent
identification. Due to a blocked N-terminus residue, it was not possible
to Edman sequence it. We would like to continue using the same sample,
now for identification by peptide mass fingerprinting. We have
experience with PMF from in gel digested samples, but never did it from
PVDF.
1) Which are the current protocols for PMF from PVDF samples?
2) Can it be done after the PVDF piece has passed through some
cycles in the sequencer?

Other question:

3) Does anyone use or know about a method for doing PMF by ES-MS,
not MALDI-MS? I don't mean LC-MS or sequencing by MS/MS; it should be
direct injection of the mixture of peptide.

Thank you.

Marcelo.

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Prof. Marcelo Valle de Sousa
Brazilian Center for Protein Research and Services - CBSP
Laboratory of Protein Chemistry and Biochemistry
Departament of Cell Biology
University of Brasilia
Brasilia - DF Brazil
CEP 70910-900

Tel: (55)(61)307-2142
Fax: (55)(61)272-4548
e-mail: mvsousa@unb.br
http://www.unb.br/cbsp
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