Say, does anybody have experience with the dual stain technique described by Dolzhanskiy, A. and Basch, R. in Journal of Immunological Methods 180 (1995) 131-140 entitled "Flow cytometric determination of apoptosis in heterogeneous cell populations"? Briefly, this technique allows for simultaneous detection of apoptotic strand breaks in the DNA of cells and cell surface markers. Cells are stained with antibodies (in my case, anti-CD3/4/8/19 conjugated to Cy-Chrome (PE-Cy5 tandem conjugates from Pharmingen)) then fixed in paraformaldehyde, postfixed in ethanol, and finally rehydrated and stained using a kit-based TdT assay (dUTP-FITC). My problem is that while both types of staining work independently, I can't get dual staining of the two simultaneously. That is, I see a nice strong FITC fluorescence from the incorporated dUTP, but no antibody fluorescence. Anybody have any idea what the problem might be? Ryan. _/ \__/ \__/ \__/ \__/ \__/ \__/ \__/rhung@vcn.bc.ca__/ \__/ \__/ \_Apoptosis=programmed cell death/ \__/ \rwhung@unixg.ubc.ca_/ \__/ \__ _/ --you can't live without it!/ \__/ \http://www.vcn.bc.ca/people/rhung \__/ \__/ \__/ \__/ \__/ \__/ \__/ \My words Copyright (C) 1997 \__
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