how to dissolve PMSF - problem in dissolving PMSF (Jul/11/2007 )

Hi..
Does anybody has idea about the solvent to dissolve the PMSF?? It is insoluble in water. I need to use PMSF in lysis buffer for protein purification.So, is there any suitable solvent that will not interfere in protein purification??

Thanks

-veerubiotech-

in absolute ethanol. I prepare 200mM stocks. good luck

-S2RNAi-

QUOTE (veerubiotech @ Jul 12 2007, 12:54 PM)

Make your stock in isopropanol. You can do 100 or 200 mM, and use at 1 mM

-swanny-

Yes, 0.1M in isopropanol (a.k.a. 100x stock)

-Almasy-

200 mM in EtOH (100%).
Refrigerate the liquid stock. Hydation kills the activity of PMSF, so the anhydrous EtOH is very important.
The stock can be stored up to 9 months @ 4C; toss it sooner if you suspect high-humidity storage.
In acqueous solutions (your RIPA or whatever you use for lysis buffer), it has a short half-life. The half-life ranges from ~110' @ pH 8 down to about a half hour @ pH 7... be sure to keep your buffer on ice if you are using it over a span of a few hours.
If you make a "lysis base" ahead of time, keep PMSF out until point-of-use, just as you would do with your enzymatic inhibitors.

-Eric

-earmstro-