I'm working with isolated PBMCs. So far, I'm using CD3, CD4, CD5, CD14, CD19, and CD20. Will this eliminate the majority of the Lin+ cells EXCEPT for NKCs? First time planning a flow cytometry experiment, and I could use the guidance of more experienced researchers. Thank you!
Hi Jaime,
if your are talking about PBMC most of the Lin+ will be eliminated.
These are the cells you will not eliminate:
myeloid DCs
Basophils
CD16+ CD14low/neg Monocytes
Your cocktail eliminates T cell, B cells (and by CD19 probably most of the CD19low Plasmablasts) and CD14+ Monocytes. I guess plasmacytoid DCs will be omitted as well (low expression of CD4).
You could try to use HLA-DR, which would help to get rid of the DCs and Monocytes, but there are a few HLADR+ NKC. Alternatively you could try CD36 which is expressed by the myeloid lineage (though expression is lower on CD16+ monocytes)
If you are talking about whole blood (RBC lysed) you should consider the neutrophils and eosinophils.
Axel
Thank you, Axel. I have clarified in my question that I'm starting with isolated PBMCs. I appreciate your input so much!
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