I need identify monocytes, macrophages and neutrophils using flow cytometry.
My antibody panel include: CD45, F4/80, Ly6C, Ly6G, CD11b.
My samples are derived from blood and bone marrow mice. Someone could me help?
Thanks!!!
Neutrophils will be Ly6G+ CD11b+. Monocytes will be mostly Ly6Chigh CD11b+, but there will also be some which are Ly6Clow. F4/80 is definitely not an exclusive macrophage marker, so you would need to use additional markers (but there wont be any macrophages in the blood). For this I recommend the following reading: http://www.nature.com/nri/journal/v14/n8/full/nri3712.html
Good luck with your experiments!
In my experience, C204 (scavenger receptor A) is the most specific marker of mouse macrophages (but may be also weakly expressed by dendritic cells, if one belives that they represent a distinct from macrophages cell type). Anti-F4/80 Ab also stains eosinophils, but combining anti-F4/80 mAb with anti-Siglec F Ab allows distinguishing macrophages (F4/80 high Siglec F neg) from eosinophils (F4/80 high Siglec F high). Anti-Ly6C Ab is completeley nonselective and labels strongly numerous types of leukocytes, so I do not know how to identify monocytes. Gr-1 (Ly6G) is a good marker of neutrophils, which are labeled strongly with anti-Gr-1 Ab. It is simpler to identify blood leukocytes on cytospine preparations stained with Giemza-May Grunwald.
- Badges
- Science method