metabolic labeling of yeast

Kevin Morano ez005528 at rocky.ucdavis.edu
Tue Feb 15 16:22:25 EST 1994


shah at RESUNIX.RI.SICKKIDS.ON.CA wrote:
: Dear people of the yeast community,
:        I would like some expert advice as to the best conditions to
: metabollicaly label yeast cells with 35S-methionine.  I would like to
: immunoprecipitate RNAP complexes from yeast cells, and knowing that they
: are not abundant proteins, I would need to label the cells really
: efficiently.  So please give me some advice as to how I can do that.  I
: thank you for your attention and am waiting anxiously for your kind replies

:                                             Sincerely

:                                             Sherry Nouraini
------------------------------
Hi Sherry, we regularly label yeast cells to look at vacuolar proteins. 
We use express-label from NEN-Dupont and label typically at .25 uCi/ul 
for 20-30 minutes. AFter various chase points, we TCA precipitate the 
proteins, extract them into immunoprecipitation buffer and go from there.

Anyway, this works really well for us. You might be able to label for 
longer periods of time (1 hour), even though its well after the linear 
range of incorporation. Good luck.
---------------------------------------------------------------------------
Kevin A. Morano
Internet:kamorano at ucdavis.edu    
Bitnet:kamorano at ucdavis   
Section of Microbiology, University of California, Davis



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