how to insure that a protein is nuclear or not?
klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Fri Feb 25 17:35:36 EST 2000
:My prot display strong NLS but immunocytochemystry failed to detect it
:in the nucleus.
:How could i cheeck whether it's because it doesn't enter the nucleus or
:because its epitope become inaccessible when in the nucleus?
If you have polyclonals, then the epitope idea is extremely unlikey and
if in immunocytochemistry your get good signal from somewhere
else but not nucleus, then it is fair to say your protein is not
nuclear. To make sure, dissolve cells in 1% triton - nuclei do not
dissolve under these conditions, retain most nuclear proteins, and
sediment at low speed. If you don't get a signal on Western from
nuclei, then you have your answer.
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