Dear colleagues Last week I asked for advice on antibodies for identification of mouse leucocyte subpopulations. Many of you have replied to tell me where I may get these - few have replied with what I really want to know, which antibodies to use for which subpopulation. My context is looking for expression of a particular cell surface molecule in different leucocyte subpopulations - and I intend to use FITC conjugate (FL1) for identification of this molecule (on a single 488nm laser cytometer), leaving FL2 and FL3 channels free for subpopulation identification. I would prefer a minimal panel of PE-conjugated antibodies (for cost and tube number limitation) to allow me to discriminate between granulocytes, monocytes, T cells, B cells and NK cells. For instance, in humans I can get a fairly good discrimination between these major leucocyte subpopulations with a panel employing CD3; CD14; CD16 and CD19 - which (with FSC/SSC and PerCP CD45 information) allows virtually complete discrimination with a 4 tube set of tests. I hope to do something as simple with mouse leucocytes - is this too optimistic? Robin Barclay SNBTS Edinburgh
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