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