We have been trying to isolate monocytes from mouse spleens using StemCell's monocyte isolation kit. However, our yields have been terrible (e.g. 200*10^6 splenocytes --> 1*10^6 macs). I heard that splenic macrophages digest RBC's and will stick to the magnetic collumn due to the ingested iron. Further, we would like all CD11b positive cells in the spleen, not just the monocytes. The reason for using negative selection as opposed to positive would be to minimize the effects of bound metallic antibodies.
We don't know of a negative selection kit, but Creative Diagnostics has a CD11b isolation system (magnetic beads) that might be of interest to you:
Our listing is here:
https://www.linscottsdirectory.com/products/creative-diagnostics/Absolute-Mag-trade-Anti-Human-Mouse-CD11b-Magnetic-Particles-10430067
You can get more details from them, by following the more info link.
I have never seen a negative selection for CD11b. I've tried the Stemcell system, which is the one that you can use just a regular tube by putting into a magnet? Yeah... those never really worked for me. I would go with Miltenyi Macs magnetic isolation kits. They really worked very well for me in the past. I can't really answer about yields, as I don't usually isolate monocytes from spleen. Mine was mostly bone marrow.
CD11b positive cells in the spleen are not very abundant and when I have tried them yield is very poor. I switched to the the bone marrow system. I plate the cells in the complete media and after 7 days you should have a very enriched and purified population. Also, I add MCSF to the media
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