Protein Extraction from Yeast

Bernard P. Murray, PhD bpmurray*STUFFER* at socrates.ucsf.edu
Wed Apr 7 17:50:51 EST 1999


In article <01be8138$b3042ea0$df019386 at schmidtdw>, "Thorsten Schmidt"
<Thorsten.Schmidt at rz.ruhr-uni-bochum.de> wrote:

> Dear reader! 
> I would like to make protein extract from a yeast overnight culture 
> to analyse it via western blot. 
> What are the easiest, fastest and satisfactory methods to do this? 
> (The methods in Sambrook et al. are frightening complicated and
> time-consuming.)
> Thank you very much in advance!!!
> Thorsten Schmidt

A better source of protocols is the Yeast chapter in
Current Protocols.  The involved/time-consuming
methods are generally for quantitative recover from
large volumes of yeast culture and cracking the cells
is the hardest part of the process.
     If you just want a small volume of soluble protein
for a western blot I would suggest beating the cells
with glass or zirconia/silica beads.  Just keep the
mix cool, buffered and include lots of protease
inhibitors.  The resulting supernatant will be crude
but should be okay for the blot.
     Have a look in the literature for some protocols
and then come back if you need more advice (you have
to show *some* sort of effort on your part).
     I would also suggest posting to bionet.molbio.yeast

     Good luck,
          Bernard
-- 
Bernard P. Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA



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