Thanks to all who replied to my original query on "nulear localization...." . Despite my typing error, I did receive some responses. The bottom line is that it still seems that the preferred method of looking for protein translocation is by fluorescent microscopy. It was suggested that the nuclear isolation method is the best bet for flow analysis. Joe Dynlacht sent me a protocol utilizing a tissue grinder which results in clean nuclear isolations in his hands. His suggestion of using an antibody to a cytoplasmic protein like beta-actin to control for cytoplasmic contamination was great and will be used. Dennis Young suggested the following reference regarding FRET to show protein co-localization: Nagy P; Bene L; Balazs M; Hyun WC; Lockett SJ; Chiang NY; Waldman F; Feuerstein BG; Damjanovich S; Szollosi J. EGF-induced redistribution of erbB2 on breast tumor cells: flow and image cytometric energy transfer measurements. Cytometry, 1998 Jun 1, 32(2):120-31. Thanks for all the responses. Julie
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