In article <firstname.lastname@example.org>,
cfankhauser at AIM.SALK.EDU (Christian Fankhauser) wrote:
> Hello everyone,
>> extracting proteins out of yeast with glass beads and hard shacking can be
> tricky, Pierce now offers an alternative with their Y-PER Yeast Protein
> Extraction Reagent, the problem is that it cost 200$ for 500ml. I know that
> it contains phosphate buffer, high salt and some ionic detergent but does
> anyone know the exact composition? or have a reference for this? or have
> any experience with it?
> Thanks for your suggestions
> Christian Fankhauser
When lysing yeast for beta-gal assays we simply add an
equal volume of 20% CHAPS. This results in complete
lysis and is (of course) compatible with beta-gal
activity (with CPRG).
Since 100 g of good quality CHAPS would cost
more than $500 I doubt if this is the composition
(unless Pierce has done a complete U-turn on its
pricing policy). Maybe a related detergent (cholate?
deoxycholate?) is the active ingredient.
Alas my protein of interest is membrane bound
so I have to stay with sonication of sphaeroplasts :-(
and I haven't bothered experimenting with other
Bernard P. Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA