Basic question about PMSF
(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.
> > 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
> Dennis (I was listening)
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