Background information
Intermediate and non-classical monocytes together are referred to as CD16+ monocytes and account for about 5–10% of all blood monocytes. CD14+ CD16+ monocytes differ from CD14+ CD16- monocytes with regard to phenotype and immunological function. CD16+ monocytes are, for example, considered more mature and are believed to have a higher T cell stimulatory capacity than CD16- monocytes.
Detailed separation procedure
Isolation of CD16+ monocytes is performed in a two-step procedure. Prior to separation, cells are incubated with FcR Blocking Reagent. Granulocytes and NK cells are then magnetically labeled with a cocktail of CD15 MicroBeads and CD56 MicroBeads, and depleted over a MACS Column. The flow-through in this first separation step contains pre-enriched, unlabeled CD16+ monocytes. In the second separation step, CD16+ monocytes are efficiently isolated by positive selection after labeling with CD16 MicroBeads.
Downstream applications
Isolated blood CD16+ monocytes are suitable, for example, for investigations on CD16+ monocyte maturation, migration, and differentiation, or for studies on antigen uptake, antigen processing and presentation to T cells.
Columns
For the first magnetic separation (depletion): LD or autoMACS® Columns. For the second magnetic separation (positive selection): MS, LS, or autoMACS Columns.
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