Hi I had heard the opposite that Ki-67 staining couldn't be done on frozen cells but our rresults showed frozen and fresh cells to be comparable. Following surface staining I used FACSlyse to permeablize the samples (and lyse the red cells in fresh samples), 15 minutes at RT followed by a couple of washes before adding the antibody. I never had much luck with anything except the Immunotech antibody. If you add a tad of Tween 20 to your wash buffer before staining and during staining the signal gets a bit brighter, about 0.05% or was it 0.5%. Don't expect more than about 3-8% positives. Hope this helps, if you need any more details let me know. Simon Monard FACS Lab Manager Trudeau Institute Saranac Lake NY12983 Ph 518 891 3080 X352
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