The difference of FITC and APC is that they are probably on the cell surface or otherwise too far from the nucleus to undergo FRET (which is limited to less than about 60 Angstroms). If one had a FITC labeled antibody in the nucleus there would be the same problem because FITC emission has strong overlap with 7-AAD emission. Ronald Rabin wrote: > I don't know why you may be having a problem with PE. I have used FITC and > APC conjugates successfully with 7-AAD and Pyronin Y. > > ron > > on 1/28/02 4:02 PM, Newsom, Brian S. at BSNEWSOM@txccc.org wrote: > > > > > Question to all of you DNA experts. We have someone who is tring to run an > > experiment staining a nuclear localized protien with PE and then staining > > with 7-AAD. The 7-AAD staining works well with a good CV. The antibody > > staining looks decent (although low percentage and fairly dim) when by > > itself, but when the 7-AAD is added the antibody staining totally goes away. > > Is there an energy transfer or steric hinderance issue that may be going on? > > Any help appreciated. > > > > Brian > > > > Ronald L. Rabin, M.D. > Senior Staff Fellow > Laboratory of Immunobiochemistry > DBPAP/OVRR > Center for Biologics Evaluation and Research > U.S. Food and Drug Administration > 29 Lincoln Drive (MSC-4555) > Building 29, Room 129 > Bethesda, MD 20892-4555 > > phone: 301.496.8806 > fax: 301.402.5177 > email: rr84g@nih.gov
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