Protease inhibitor coctail for yeast?

Rob Kirkpatrick kirkpat at cc.?umanitoba?.ca
Fri Oct 31 10:57:10 EST 1997


In article
<Fergus.Doherty-ya02408000R2910971649140001 at news.nottingham.ac.uk>,
Fergus.Doherty at nottingham.ac.uk (Fergus Doherty) wrote:

> I seem to be getting quite a lot of protein degradation in yeast
> homogenates and subfractions (glass beads to break cells), even using PMSF,
> leupeptin and pepstatin.  Sigma cell a cocktail of AEBSF (like PMSF?),
> E-64, pepstatin A and o-phenanthroline (metal chelator?).  Is this any
> good, and suggestions for a good cocktail.  If possible I would like to
> leave out a calcium chelator - would this cause a problem?
> 
> Thanks
> 

I don't have a complete answer for you, but check out Boehringer
Mannheim's web page - they have a section on protease inhibitors including
cocktail "tablets".  I haven't used these so I can't speak for their
usefulness but maybe someone else can.  I use a mix of Pepstatin A,
Leupeptin, Chymostatin, Aprotinin, Antipain and PMSF.  I'm about to add
Bestatin to that list. (All from SIGMA)  I still get some degradation, but
the stuff I want to see is definitely there.  My list sounds like
overkill, but better safe than sorry.  Here's the web site address for the
protease inhibitor page:
http://biochem.boehringer-mannheim.com/prod_inf/manuals/protease/Prot_cht.htm

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