Basic question about PMSF
Thu Dec 14 17:47:46 EST 2006
On 2006-12-14 17:42:43 -0500, Dennis said:
> On 2006-12-13 12:03:47 -0500, "Breslauer" <breslauer1981 At gmail.com> said:
>> 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?).
>> Thans for any clue
> 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
OOps, I was thinking of TPCK. Yes, PMSF would be useful to inhibit
proteases, but it is a rather nondiscriminate alkylator and could cause
More information about the Methods