I want to use FACS sorter to sort/isolate monocytes from human BPMCs and based on literature CD14 is the best marker. After sorting Monocytes, I need the CD14 to be available/free for LPS and probiotics stimulation studies. Could anyone advise me if I use Anti CD14 conjugated Ab for sorting Monocytes, how can I get rid of CD14 bindings and how I can release/free CD14 for further LPS and bacterial stimulation/induction?
I have been advised the FACS Ab will be bound to part of CD14 and not all of it, and I can still use CD14 for stimulation. I haven't found any papers yet. Does anyone have any experience about this please?
What's the best sorting protocol to maximise the yield and the viability of the monocytes?
Your help is very much appreciated.
Thanks many