CD16+ Monocyte Isolation Kit
Description The CD16+ Monocyte Isolation Kit has been developed for the isolation of the CD16+ monocyte subset from PBMCs, which accounts 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, supposed to be more mature and have a higher T cell stimulatory capacity than CD16- monocytes.1,2 Isolation of CD16+ monocytes is performed in a two-step procedure. First, granulocytes (neutrophils and eosinophils) and NK cells are magnetically labeled and depleted using a cocktail of CD15 and CD56 MicroBeads. Neutrophilic granulocytes and NK cells have to be depleted in advance, because they also express CD16. Thereafter, the CD16+ monocytes in the non-magnetic flow-through fraction are specifically labeled with CD16 MicroBeads and efficiently isolated by positive selection. 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.		Figure 1 CD16+ monocytes were isolated from PBMCs using the CD16+ Monocyte Isolation Kit. In a first step, granulocytes and NK cells were depleted. In a second step, CD16+ cells were enriched. An LD Column and a MidiMACS™ Separator were used for the depletion step, and an MS Column and a MiniMACS™ Separator for the positive selection step. Aliquots of the different cell fractions were stained for CD16 and CD15/CD56 or CD14, respectively. PBMCs before separation Pre-enriched CD16+ monocytes Enriched CD16+ monocytes Figure 2 Working scheme for the isolation of CD16+ monocytes from PBMCs.