Dear Flowers, Does anyone have any experience using an intracellular flow assay for phosphorylated nuclear proteins? I am interested in detecting phosphorylated STAT transcription factors, which are nuclear proteins and pose the additional challenge of forming dimers between their phospho-epitopes and SH2 domains. I have been using a method based on PFA fixation and MeOH permeabilization, but I have encountered problems, which, I think, are related to nuclear permeabilization. Any suggestions would be greatly appreciated. Thanks, Traci --- Traci Battle, Ph.D. Deparment of Adult Oncology Dana-Farber Cancer Institute, M549 44 Binney St. Boston, MA 02115 Tel (617)632-5397 Fax (617)632-5167
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