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PMSF dire straits

pathos pathos at freerealtime.com
Sun Jan 30 13:57:17 EST 2000


In article <86tjvs$81$1 at bgtnsc03.worldnet.att.net>, "simm"
<simm at worldnet.att.net> wrote:

> I am currently running experiments requiring the use of PMSF. I have to
> prepare fresh
> solutions each time which as you may know takes up time and wastes a lot of
> the
> reagent. Is there a way to prepare a solution which is stable in storage?
> are there other similar protease inhibitors? I am specifically interested in
> stability of PMSF in methanol,  acetone and isopropanol over time. I have
> heard some colleagues store such solutions for up to a month, but I need
> confirmation from the scientific body.

The problem with PMSF is not the stability in ethanol or methanol which is
years, it is the stability after you place it in agueous media (like lysis
buffer).  The half life now is about 20 minutes.  For my purposes, that
stinks.  Since I do not like to use PMSF, I have found a substitute which
is more stable in aqueous solution and not as toxic as PMSF.  The
substitute that I use is AEBSF.  The protease that I work with or at least
am concerned with is uPA.  Even at -20, I can see some of the effects of
its activity. 

regards,

-- 
Peter Pediaditakis
University of Pittsburgh
Dept. of Pathology




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