Hi, OK so we use a FACS Vantage SE. The filter sets for the He/Ne laser are according to config. 6 in the BD Immunocytometry product booklet, i.e.: on FL4 the 780/60 filter for APC-Cy7 on FL5 the 660/20 filter for APC the 710 LP inbetween As for your second question, changing to PE may be purely informative (steric hindrance issue), but it's excluded in this experimental setting (we're screening a whole battery of other markers in PE in these 5-color experiments) Hope this clarifies the question Tx again Karim "Fischer, Randy (NIAMS)" wrote: > Karim, > > I guess the first question I have for you is what instrument are you using. > Next, what are the filter sets/dichroics. Finally, have you tried different > fluorochrome combinations to eliminate steric hindrance of the antibodies > (perhaps changing one to a PE conjugate since this is also a fairly large > protein). > > Randy T. Fischer > NIH/NIAMS > Autoimmunity Branch > Building 10, Rm 6D57 > 9000 Rockville Pike > Bethesda, MD 20892 > (301) 594-3537 > fischer1@mail.nih.gov
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