EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion

Protocol tips

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		Remaining cells were stained for FACS with anti-CD14, -16, -HLA-DR, and lineage exclusion markers as described for phenotyping above.

Downstream tips
Remaining cells were stained for FACS with anti-CD14, -16, -HLA-DR, and lineage exclusion markers as described for phenotyping above.

Publication protocol

Leukocyte concentrates were purchased from the Central Blood Bank of Pittsburgh. Red blood cells (RBC) were lysed using an ammonium chloride buffer (Stem Cell Technologies) according to the manufacturer’s protocol. PBMCs were rinsed in wash buffer twice, then incubated at 37°C in FCS containing 5mM EDTA to remove any remaining platelets [30,31]. After washing, monocytes were negatively selected using an EasySep Human Monocyte Enrichment Kit without CD16 Depletion (Stem Cell Technologies) according to the manufacturer’s protocol. Remaining cells were stained for FACS with anti-CD14, -16, -HLA-DR, and lineage exclusion markers as described for phenotyping above. High-speed sorting was performed on a BD FACSAria to yield high-purity monocyte subset populations (>95%). Cells were cultured in complete medium consisting of RPMI-1640 (Cellgro) supplemented with 2 mML-glutamine, 10mM HEPES, 100 IU/mL penicillin/streptomycin and 10% heat-inactivated fetal calf serum (FCS) (all from Gibco) except where other media formulations are indicated

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Manufacturer protocol

Download the product protocol from STEMCELL technologies for EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion below.

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