internal peptide sequencing

Peter Hunziker phunzi at bioc.unizh.ch
Wed Aug 26 01:48:33 EST 1998


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



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Dr. Peter Hunziker
Universitaet Zuerich, Biochemisches Institut
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CH-8057 Zuerich, Switzerland

Tel: 01/365 55 20
Fax: 01/365 68 05
E-mail: phunzi at bioc.unizh.ch

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