Basic question about PMSF

Pow Joshi via methods%40net.bio.net (by pow.joshi At gmail.com)
Fri Dec 15 12:14:01 EST 2006


On 12/14/06, Dennis At oat.bio.indiana.edu <Dennis At oat.bio.indiana.edu> wrote:
> On 2006-12-13 12:03:47 -0500, "Breslauer" <breslauer1981 At gmail.com> said:
>
> > Listen,
> > is it normal that when adding PMSF (250mM solution in MeOH) to cell
> > free extract from E.coli (broked by sonication and spinned 12000g for
> > 30min) to the final concentration of 0,5mM buffer becomes cloudly and
> > st become to precipitate (probably protein?).

I wonder if it is because your PMSF is in methanol. You could try
using ethanol solubilised PMSF.

cheers
pow
> >
> > Thans for any clue
> >
> > Julio
>
> You are alkylating at least some protein and increasing its
> hydrophobicity, so a precipitate is not surprising.
>
> Why on earth are you adding PMSF to a protein extract?  The common use
> of PMSF is to add it to a water solution to alkylate and inactivate
> RNAses.
>
> Dennis (I was listening)
>
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