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USA::RGB12955 RGB12955%USA.decnet at USAV01.GLAXO.COM
Wed Aug 3 17:49:29 EST 1994


For a very rapid look at yeast proteins, try the method in Wright et al,
Yeast 5:51-53 (1989).  Basically, pipette 10e7 - 10e8 cells into ufuge
tube, add TCA to 10% final (v/v) on ice.  Leave on ice (I use 20min).  Spin
10min in cold, wash 2X in 100% EtOH with 10min on ice each time.  Vacuum
dry, boil in 2X sample buffer, load, run, stain or blot, etc.  Blot time is
about twice normal, as "proteins that have been TCA precipitated appear to
transfer more slowly" according to the authors.  Oh!  don't forget to spin
after boiling to remove residual cell debris.  10e7 cells per lane is plenty
for Coomassie stain.

Rich Buckholz
RGB12955 at usav01.glaxo.com




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