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Re: MAP kinase translocation
Observing good nuclear localization is dependent on cell type (some cells
don't translocate a very high percentage of the total protein), and on the
timing. You should stimulate your cells, then do a time course. Sometimes
the "window" of good nuclear localization can be narrow.
You may also need to try different fixation conditions as these proteins
are small enough to diffuse back out of the nucleus if not adequately