[Protein-analysis] Re: Candida proteomics
pDOTpagel at gsf.de
Mon Mar 13 03:49:26 EST 2006
jaya at hkusua.hku.hk wrote:
> I want to extract proteins from Candida biofilms grown in 1cmm acrylic coupons.
> Obtaining cell pellet is pretty small although I pool few coupons together.
> Can you please tell me a protocol to extract proteins from this. Here I am
> looking for extrac-cellular proteins as well as Candida cellular proteins.
> I tried with glassbeads, it didn't work.
I have never done that with Candida - but for S. cerevisiae the glass
bead method worked well for me. A few things to consider: Make sure you
have the right size beads - if they are too large grinding doesn't work.
You have to vortex *vigorously* and long enough. Try a few different
lysis buffers - I had good results with an SDS/urea buffer. Maybe try to
use more cells.
Clontech has a pretty good protocols handbook online - it's for S.
cerevisiae but I think most of the recipes should work just as well for
hope it helps
Dr. Philipp Pagel Tel. +49-8161-71 2131
Dept. of Genome Oriented Bioinformatics Fax. +49-8161-71 2186
Technical University of Munich
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