I would like to do FACS sorting for mouse monocytes after ficoll separation. However, I am confused for information I get about those surface markers for mouse monocytes. Some listed about 18 abs conjugated with flurochromes. But some listed 4 to 5 only. So what are the most commonly essential Abs for FACS sorting of mouse monocytes.
Do you mean you want to isolate monocytes for further experiments? You don't need a sorter, those are usually for unusual cell types. You can use any of the commercial kits for mouse monocyte isolation.
If you want to do flow analysis on monocytes, the easiest is to gate for CD11b+GR1-, or you can actually just use FSC/SSC (monocytes are pretty well separated from lymphocytes and granulocytes) or you can also try CD11b+MHCII+.
Hi Ming Chuan:
This issue has been discussed before in Resarchgate, which should be helpful. https://www.researchgate.net/post/Whats_the_best_maker_for_monocyte_macrophage_in_mouse_peripheral_blood.
Two main methods for monocytes purification: sorting and beads separation. Sorting produces higher purity, but the sorted cells stressed by high pressure, may be contaminated during sorting; Beads separating is easier to manipulate, won't affect cell function (especially negative selection), less chance to be contaminated; but lower purity, higher budget. The choice depend on your experimental aims, your budget, expertise et al.
Regards
Shiqiu
Hi Shiqiu, I found it before. Still so many suggestions and they were varied one to another too and finally very confusing. This is why I raised this again. thanks
Monocytes are cd11b+/ly6g-. There is often confusion with ly6c, ly6g-, and gr1. Ly6g is expressed on granulocytes, ly6c is uniformly expressed on granulocytes (intermediate) and variably expressed on monocytes (hi low neg), and gr1 is a shared epitope between ly6c&g. Ly6c indicates monocyte approximate age and phenotype. Ly6c hi = young ccr2 hi / cx3cr1 low cells and ly6c lo = older ccr2 lo / cx3cr1 hi. These equate to classical and non classical cells. There are also cd115 f4/80 and some other markers but they aren't necessary for identifying monocytes in mouse blood.
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