In article <01HQ3XX4F53SBLD26P at PORTIA.CALTECH.EDU> EARN at PORTIA.CALTECH.EDU (JUST ERIC) writes:
> I'm looking for help in creating an effective protease inhibitor
>cocktail for yeast extract - I am expressing a small soluble enzyme in
snip
>> I have also considered adding EDTA
Keep EDTA in there everywhere you can. I had enormous proteolysis problems
with one prep, where I tried to avoid EDTA for the same reasons that you
did. All my problems disappeared when I added EDTA back. The other thing
to do is to make all your buffers metal free by putting them through
chelex and washing all bottles and glassware etc with chelex water. Personally
I'd rather work in a space suit myself.
Joe Mack mack at ncifcrf.gov
