MagniSort™ Human pan-Monocyte Enrichment Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Briefly, a single-cell suspension containing 1 × 108 PBMCs per mL of cell separation buffer (PBS, 3% FBS, 10mM EDTA) was prepared.	Cells were then incubated for 10 min with 20 μL of MagniSort™ enrichment antibody cocktail per 100 μL of cell suspension. Cells were washed and resuspended in separation buffer. Then 10 μL of MagniSort™ negative selection beads were added per 100 μL of cell suspension. Following 5 min of incubation, a magnet was used to remove the bead-bound non-monocyte lymphocytes from the monocytes.	. Enrichment efficiency was determined by flow cytometry staining with anti-human CD14 eFluor 450 (clone 613D, eBioscience) and anti-human CD16 APC (clone CB16, eBioscience).

Upstream tips
Briefly, a single-cell suspension containing 1 × 108 PBMCs per mL of cell separation buffer (PBS, 3% FBS, 10mM EDTA) was prepared.
Protocol tips
Cells were then incubated for 10 min with 20 μL of MagniSort™ enrichment antibody cocktail per 100 μL of cell suspension. Cells were washed and resuspended in separation buffer. Then 10 μL of MagniSort™ negative selection beads were added per 100 μL of cell suspension. Following 5 min of incubation, a magnet was used to remove the bead-bound non-monocyte lymphocytes from the monocytes.
Downstream tips
. Enrichment efficiency was determined by flow cytometry staining with anti-human CD14 eFluor 450 (clone 613D, eBioscience) and anti-human CD16 APC (clone CB16, eBioscience).

Publication protocol

Monocytes were isolated from thawed PBMCs using the MagniSort™ human pan-monocyte enrichment kit (eBioscience). Briefly, a single-cell suspension containing 1 × 108 PBMCs per mL of cell separation buffer (PBS, 3% FBS, 10mM EDTA) was prepared. Cells were then incubated for 10 min with 20 μL of MagniSort™ enrichment antibody cocktail per 100 μL of cell suspension. Cells were washed and resuspended in separation buffer. Then 10 μL of MagniSort™ negative selection beads were added per 100 μL of cell suspension. Following 5 min of incubation, a magnet was used to remove the bead-bound non-monocyte lymphocytes from the monocytes. Enrichment efficiency was determined by flow cytometry staining with anti-human CD14 eFluor 450 (clone 613D, eBioscience) and anti-human CD16 APC (clone CB16, eBioscience).

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