[Protein-analysis] Re: Candida proteomics

Philipp Pagel 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
Candida:

http://www.clontech.com/clontech/techinfo/manuals/PDF/PT3024-1.pdf

hope it helps

	Philipp


-- 
Dr. Philipp Pagel                          Tel. +49-8161-71 2131
Dept. of Genome Oriented Bioinformatics    Fax. +49-8161-71 2186
Technical University of Munich
http://mips.gsf.de/staff/pagel


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