Maciej Simm wrote: > > One of the people I'm working with right now is studying apoptosis in > periteneal macrophages using a kit with annexin V monoclonal FITC ab > and propidium iodine. > > The cells are autofluorescent. The "unstained" tube has signal in up > to 2nd log. > If you're having trouble with high autofluroescence, you';ll do well to move away from the green and into the red part of the spectrum. Annexin APC would be a better choice for example. Or an alternative assay may be more appropriate: TUNEL biotin with a red avidin. As an aside, we recently observed very high levels of constitutive annexin binding to freshly isolated monocytes. We didn't follow up on this as we were interested in lymphocyte death and are not sure of its significanece. I strongly recommend using DNA staining and fluorescence microscopy to verify you r flow assay for apoptosis. Good luck, Tom ***************************************************************************** Thomas W. Mc Closkey, Ph. D., Director of Flow Cytometry North Shore University Hospital, New York University School of Medicine Boas Marks Biomedical Research Center, 350 Community Drive Manhasset, Long Island, New York 11030 ph: 516-562-4844 [office], 516-562-1135/4641 [lab] fax: 516-562-2866 *****************************************************************************
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