F4/80 is used as a marker to identify murine tissue/peritoneal macrophages.
In our working groop we isolate monocytes from rat tibia and femur and differentiate them subsequently into bone marrow-derived macrophages by using M-CSF. As there is no anti-rat F4/80 antibody for FC application, we use murine antibodies instead. Yet using those antibodies failed to give us a valid signal for both FC as well as IHC applications. In other working groups the same antibody worked for rat tissue macrophages, which led us to the question if M-CSF macrophages show different F4/80 expression pattern compared to tissue macrophages?
Does anyone has experience with this? Or maybe with confirmed rat-mouse crossreactivity of F4/80 antibodies?
Thank you in advance!
yes they do. I used it for murine bone marrow-derived cells. M-CSF differentiated expressed higher F4/80 compared to GM-CSF differentiated cells. I used flow cytometry to measure the expression. Thank you
I have used M-CSF and 30% L929 conditioned medium to differentiate murine bone marrow into macrophages. Then used F4/80 and CD11b as markers to identify macrophages. Both of them give a as high as 96% F480+CD11b+ percentage. Check if this antibody works for mouse or not, check if M-CSF expired. Then try to differentiate BMs for longer time (7days-9days) or use higher con. of M-CSF
Yes, they express the markers. From our experience, we always check beyond differentiation day 7-9. The majority of them (80-90%) will express F480
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