You may try to digest the protein in the gel and analyze the fragments by LC/MS
or NanoSpray-MS and MS/MS or by MALDI-TOF-MS. Perhaps your colleagues from the
Syn/Seq Facility in the Molecular Biology Dept. may help you. Their URL is:
http://www.molbio.princeton.edu/synseq/synseq.html
I do not know the lab but in the ABRF (http://www.abrf.org) Yellow Pages
(http://www.abrf.org/cgi-bin/ysearch.cgi) they offer sequencing and MS services.
best regards
Peter
kraepiel wrote:
> Some time ago there was a discussion about the best way to go about
> digesting a protein to obtain internal peptide sequence. Of course I was
> not paying attention then and now I need the information.
> I have a partially purified protein preparation and can with which I
> can get an isolated band of my protein of interest by SDS PAGE. If I
> electroelute this protein, digest it and run a gel to obtain isolated
> fragments I will probably lose a lot during these manipulations. This
> protein is pretty low copy number and it is difficult to get enough to
> sequnce. I would be grateful for any alternative strategies one might
> use to get internal sequence.
>> Todd Lane
>Tlane at princeton.edu
--
************************************************************
Dr. Peter Hunziker
Universitaet Zuerich, Biochemisches Institut
Winterthurerstr. 190
CH-8057 Zuerich, Switzerland
Tel: 01/365 55 20
Fax: 01/365 68 05
E-mail: phunzi at bioc.unizh.ch
************************************************************
Visut our department home page: http://www.unizh.ch/biochem/
************************************************************
