Dr E. Buxbaum
EB15 at le.ac.uk
Mon May 11 18:55:32 EST 1998
François Mallet wrote:
> We try to purify a protein from human urine. The goal of our our work is
> to sequence this protein that have an biological activity. However, after
> multiple steps of purification, and although the specific activity of the
> enriched protein increased, this protein remains contaminated with large
> amounts of albumin. Thus we are unable to sequence it.
How do you tell that there is albumine contamination? The method which
gives you that information can also be used to purify the albumine away.
Remeber that for sequencing purpose you need inly a few hundred picomole
of your protein, that much can be isolated for example by SDS-PAGE and
blotting onto PVDF membranes. Stain with Coomassie the usual way, cut
out the bands and give the snippets to the sequencing laboratory. The
PVDF serves as a carrier during the sequencing process.
If you need more help, than you have to give us more info on your
More information about the Proteins