Ruedi, Foxp3 protein levels should be the same or greater after stimulation, however, we have found that the cytokine permeabilization conditions do not achieve very good Foxp3 staining, while the Foxp3 permeabilization protocol does not preserve good cytokine staining. I wonder if anyone out there has tried a hybrid protocol, which gives reasonable nuclear and cytoplasmic staining for both these antigenic targets... AW Andrew D. Wells, Ph.D. Assistant Professor of Pathology and Laboratory Medicine University of Pennsylvania School of Medicine Joseph Stokes, Jr. Research Institute Biesecker Pediatric Liver Disease Center The Children's Hospital of Philadelphia 916E Abramson Research Center 3615 Civic Center Boulevard Philadelphia, PA 19104 phone: (215) 590-8710 fax: (215) 590-7384 On Dec 20, 2007, at 12:59 PM, Rudolf Braun wrote: > Hi, > > the staining for intracellular cytokines includes a 4 hour > incubation at 37C with > brefeldin A, PMA, and ionophore. Does this activation change the > level of FoxP3 detected > by intracellular staining ? > > Kind regards and Merry Christmas > > Ruedi > > > > > ---------------------------------------------------- > Rudolf K Braun, Ph.D. > Assistant Professor > Thoracic CV Surgery > Loyola University Medical Center > Bldg: 101; Rm: 2711 > 2160 South First Avenue > Maywood, IL 60153 >Received on Sun Dec 23 17:58:00 2007
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