Looking to isolate mouse monocytes (not macrophages) from the blood in mice.
I know people use Ficoll + MACS columns but i just read that they might activate the monocytes a bit. My lab is considering using the EasySep magnet and isolation method. Does anyone use this method and is it worth the cost?
Im trying to find the best way for us to isolate monocytes from the blood without changing the cells themselves.
1-One way is to use FACS cell sorting. It would provide you with higher purity.
You can gate on CD11b+ Ly6G-
And maybe include Ly6C+ for classical monocytes, and Ly6Clo for patrolling monocytes.
2- Another method is to use Monocyte Isolation kit from StemCell
Catalog # 19861
Also, to obtain higher numbers of monocytes, I would also recommend isolating from the Bone Marrow as well.
Best of luck with your experiment
Just to clear something up for you, macrophages don't exist in circulation, so any monocyte lineage cells from circulation are monocytes.
I agree with the previous responses that you have essentially two options for isolating circulating monocytes: FACSorting and magnetic isolation. If you want to avoid labeling your monocytes with antibodies, you can use florescent reporter mice with FACS.
It would be helpful to know what the context of the experiment and assay are to give a better answer.
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