hi everybody i'm using the boehringer-mannheim BrdU-FLUOS kit to look at GM-CSF induced cell prolif. the problem i'm having is that the FL1 fluorescence (anti-BrdU-FLOUS) is highly variable between duplicate tubes. on a FACS, i'm having to change the FL1 PMT volts by over 200 to get populations to fall in the same gate. PI staining seems OK. FL3 populations fall in similar channels without changing settings. i know that the Ab-Ag dissociation rate changes with volume of cell suspension and i've tried to be consistent with all tubes. if anyone has good experience with this product or has overcome similar problems, i would appreciate the feedback. thanks and just flow baby gary woo
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