Thanks for all the suggestions! Any more antibodies and we will blow our budget out of the water!! We have ordered NK and T-cell markers for a start. Summary: To characterize B220+IgM- population in our knockout mice (immature B cells?): 1) Ly-6D 2) CD25 3) IgM+/IgD+ for mature B cells 4) compare intensities of B220 (bright for mature/dim for immature) 5) Kappa/Lambda to rule out T-cell possibility 6) CD43, 6C3(BP1) and IgM to quantify % Pro-B, % Pre-B, and immature/mature B cells 7) CD19 as a marker for B lineage, check to see if it is CD3+ 8) B220 is expressed on activated NK and T cells (LAK cells) as well as BM progenitor cells of the NK lineage. If immature B cells, look at CD43, BP-1, and CD24. If CD19- (i.e. non-B lineage), relevant T and NK markers such as CD3, DX5, PK136, CD94 may be the appropriate markers to look for. 9) NK cell markers as well, such as NK1.1 or DX5, to look for expansion of NK cells in your mice. Many, if not all, NK cells, at least in B6 mice, coexpress B220, albeit to lower levels than B cells. Derek Schulze Cancer Research Labs Queen's University 353 Botterell Hall Kingston, ON, K7L 3N6 (613) 533-6635 http://meds.queensu.ca/medicine/crl/
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