> Xinjian Chen M.D., Ph.D." <xchen2@emory.edu> - 4/11/02 9:00 PM > > Greetings to everybody > I have recently obtained some intriguing flow data, as described below, and > am looking for help to understand its nature. When I perform multi-color > staining on spleen or lymph node cells of immunized mice with anti-CD4-FITC > plus other antibodies, I can unequivocally identify by flow cytometry a > subset (fifteen to 30%) of CD4+ T cells that are positive for B cell > markers, including B200, IgM, IgD, MHC Class II, etc. However, if I use same > anti-CD4 antibody conjugated to other flurochrome, such as PE or PerCP, than > FITC for multi-color staining, the B cell marker-positive subset of CD4 T > cells becomes no longer identifiable. > Your comments will be greatly appreciated. > > > Best regards > Xinjian Hello Xinjian, One possibility, the anti-CD4-FITC is contaminated with some "anti-mouse B marker-FITC" (eg. aCD19-FITC). An unchanged pipette tip could be the reason. Try a new batch of anti-CD4-FITC. Best regards, Gogy Peter Gogolak, Ph.D. Institute of Immunology University of Debrecen, Hungary Phone/Fax: +36 52 417159
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