Adrian, with the DAPI staining seems strange to me. That GFP staining is whiped out by HCL treatment, is o.k., because HCL extracts proteins. Perhaps you can use another detection method, how long did you label the cells and what was the concentration of BrdUrd during incubation? Depending on that, you may be able to detect proliferation by using a combination of PI and TO-PRO-3, excited with a HE-Ne laser and a 488nm laser. Best regards from Munich Wolfgang GSF- Flow cytometry group Ingolstaedter Landstr. 1 85764 Munich, Germany > -----Ursprüngliche Nachricht----- > Von: Senderowicz, Adrian (NCI) [mailto:sendero@helix.nih.gov] > Gesendet am: Montag, 14. Juni 1999 18:02 > An: Cytometry Mailing List > Betreff: re: BRDU DNAse > > > Dear flowers, > I am transfecting cells with GFP and when I try to do BRDU > Immunefluouresnce > with the HCl protocol, all DAPI and GFP staining dissappears... I > tried twice > the DNase method without any luck. Can you help? > thanks a lot > > Adrian Senderowicz, M.D. > DTP Clinical Trials Unit > National Cancer Institute > 10 Center Dr. MSC 1594 > Bldg 10, rm 6N113 > Bethesda, MD 20892 > 301-496-4119 > 301-480-7456(fax) > sendero@helix.nih.gov > > >
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