Dear Fred,
I am sending you some references for non-SDS protein extraction
from the gel. They are quit simple, compatible with MS and they should work
with 25 kDa protein, I guess. I would nevertheless try to adjust the
protocols and to look for the best suitable conditions.
1. H. Ehring et.al. Rapid Comm. in Mass. Spectrometry (1997),11,
1867-1873.
2. S. Kohen, B.Chait. Anal. Biochem. (1977), May,1;247(2):257-267.
It was also a poster on ASMS (I should look into my notes)
declaired non-SDS extraction of proteins up to 110 kDa.
There are, of course, other methods of elution and extraction not
easily compatible with MS and normally followed by different clean-up
procedures. I can also provide you with some more, in case you are
interested in.
I, personally, have tried different methods and their modifications
to extract 7.25 kDa hydrophobic protein from SDS PAGE. I attach you the
only procedure which work in this particular case.
Good luck, do not hesitate to contact me,
Alla
--============_-1275935858==_============
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--============_-1275935858==_============
Content-Type: text/plain; charset="us-ascii"
Alla Shainskaya, Ph.D
Head of Mass Spectrometry Unit,
Department of Biological Services,
Weizmann Institute of Science, Rehovot,
76100, Israel.
tel: 972-8-9343113
fax: 972-8-9344113
--============_-1275935858==_============--