yeast extraction using PIERCE Y-PER

Bernard P. Murray, PhD bpmurray*STUFFER* at socrates.ucsf.edu
Mon Mar 1 20:00:02 EST 1999


In article <l03010d00b300c6173175@[192.31.153.170]>,
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
detergents.
     Bernard
-- 
Bernard P. Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA



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