Great thanks to all who sent me information on the markers and their levels of expression activation. (1) Here is an amazing website on monos/macs: http://www.path.ox.ac.uk/sg/ (2) In general, here are the markers present on this population of cells that are USEFUL for surface staining: CD11b--medium bright on monocytes and neutrophils. CD11c--present on monocytes; easier to see in higher side scatter than T/B cells. CD13--medium bright on monocytes and neutrophils. CD14--stains monocytes, but is definitely shed on activation or when held in culture. CD15--brightly stains neutrophils. CD33--monocytes stain bright, granulocytes stain dim. CD64--present constitutively on monocytes and is not shed on activation. Be careful though because CD64 becomes upregulated on activated neutrophils. F4/80--Serotec lists their antibody against F4/80 antigen as clone CI:A3-1 (the original, I believe) Caltag has a clone called F4/80. (3) Specifically to gate on monocytes/macrophages, Mark Edinger of B-D suggests that we use CD64 and CD33. In fact we could use CD33 with CD45 and side scatter to pull out the monos, or just CD33 and side scatter alone. The huge benefit from this method is to use the complexity of the monos/macs to get rid of background staining in other, smaller cells. I have had MUCH cleaner data, personally, since using this method. Thanks Mark. Special thanks to Keith Bahjat (U.Florida College of Med.), Jan Nicholson (CDC), Abby Kelliher (Mass General Hospital), and the ever-helpful Mark Edinger (B-D, Pharmingen). Happy flow-ing... JDT ********************* Julie Davis Turner Postdoctoral fellow Centers for Disease Control & Prevention NCID/DASTLR/Tb Atlanta, Georgia USA
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