Julie We only started looking at antiphosphotyrosine by flow a few weeks ago and are using clone 4G10 biotinylated from Upstate. We have used the same standard paraformaldehyde fixation and saponin permeabilization procedure that we use for most other intracellular markers. We have been able to detect increases in mAb staining in human T cells that have been activated by crosslinking CD3. We have not been able to detect any increases when using gentler methods of activation. Barbara J Taylor Stem Cell Lab Children's National Medical Center Washington DC
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