This may be elementary to some, but it is new to me. I am
trying to extract the nuclear and cytosolic protein fractions from yeast (S.
cerevisiae). I prepared spheroplasts, dounced them and recovered the nuclei by
spinning down the spheroplast lysate at 20,000g. Is this an appropriate speed
for nuclei? Also, I have been using Nsp1 (Nulcear surface porin) as a marker
for the nuclear fraction and am facing some problems with that. What other nuclear
markers are commonly used for Western blotting? Any input regarding the speed
for nuclei spin down and the nuclear marker will be most welcome.
Thanks!
Anupama Shanmuganathan
Graduate student
Georgia State
University
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