Hi Eitan,
As always the answer depends on the questions you want to answer :-) Do you want to distinguish AML and ALL or subtypes thereof?
I am mostly working on murine AML models and interested in the maturation stage of the AML cells. What I usually do is a "normal" myeloid panel that includes Mac-1, Gr1, c-Kit and sometimes Sca1 (bulk AML cells will often not express Sca1). You could also add F4/80 to identify monocyte/macrophage-like cells.
Going deeper into AML cell phenotypes would mean to include CD34 and CD16/32 to see if you AML cells correspond to a certain myeloid progenitor type.
If you want to look at AML and ALL, you might want to include CD19 (or B220) and CD3 (or a CD4/8 combo) to distinguish between B- and T-ALL.
So, without knowing more about the questions you want to address: I would probably start with Mac1/Gr1/CD3/CD19/c-Kit and go from there...
Hope that helps,
Chris
Hi Christian, I've pretty much come out eventually with the same protocol.
Thanks a lot for your help.
Did you try CD45xSSC to separate the myeloid gate from the blast gate-then go from there?
Yes I did, strangely, most of the leukemic blasts were CD45 negative. could be a problem with the antibody. Thanks
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