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| CD16+ Monocyte Isolation Kit |
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| Overview |
| The CD16+ Monocyte Isolation Kit has been developed for a two-step isolation of the CD14+CD16+ monocyte subset from PBMCs. |
| Details |
Background information
Intermediate and non-classical monocytes together are referred to as CD16+ monocytes1 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.2, 3
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. |
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| 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. |
| PBMCs before separation |
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| Pre-enriched CD16+ monocytes |
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| Enriched CD16+ monocytes |
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| Figure 2 |
| Working scheme for the isolation of CD16+ monocytes from PBMCs. |
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| Details |
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| Products |
| CD16+ Monocyte Isolation Kit, human |
- for 2×109 total cells
Download datasheet
130-091-765
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| References |
| 1. Ziegler-Heitbrock et al. (2010) Blood 116: e74–80. |
| 2. Ziegler-Heitbrock et al. (1996) Immunol. Today 17: 424-428. |
| 3. Grage-Griebnow et al. (2001) J. Leukoc. Biol. 69: 11-20. |
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