From: Deborah Berglund (umbbd@gemini.oscs.montana.edu)
Date: Sat Dec 09 1995 - 16:48:48 EST
Try getting your antibody conjugated to biotin, then use the biotin avidin linkage with a known number of fluorescein (or PE...) molecules. Worked for us. Deb from Bozeman, MT On Thu, 7 Dec 1995, Susan & Ulrik Sprogxe-Jakobsen wrote: > Hello out there! > > I am performing flow cytometry analysis on human red cells. Due to the low > number of antigen copies per cell, the resulting fluorescense (FITC) is > close to the flowcytometer (FACScan) noise level. > > 1. Does anyone know how to calculate the real number of cell-bound FITC > molecules (and hence antigens), when working in this low area of > fluorescence intensity? > > 2. Unmarked human red cells have a lower green (FITC) autofluorescence > than certified blank beads. Any suggestions on how to incorporate this in > the calculations of cell-bound FITC molecules on human RBCs? > > > Any suggestion appreciated ... > > Best regards > > Ulrik Sprogoe-Jakobsen > > Dept. Clinical Immunology > Odense University Hospital > DK-5000 Odense C > Phone: +45 6541 3576 > Fax: +45 6612 7975 > > >
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