EasySep™ Human Monocyte Enrichment Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		Unwanted cells are labelled for removal with bispecific tetrameric antibody complexes recognizing the respective target antigen (CD2, CD3, CD16, CD19, CD20, CD56, CD66b, CD123, or glycophorin A) and dextran-coated magnetic particles.

Downstream tips
Unwanted cells are labelled for removal with bispecific tetrameric antibody complexes recognizing the respective target antigen (CD2, CD3, CD16, CD19, CD20, CD56, CD66b, CD123, or glycophorin A) and dextran-coated magnetic particles.

Publication protocol

For the analysis of LO activities, neutrophils and monocytes were routinely isolated from human peripheral blood of healthy adult male and female donors according to reported methods13,14 (for details of monocyte isolation via adherence, see Supplementary Materials online) or by positive and negative immunomagnetic selection, respectively (applicable for Figure 5E and F). For positive selection we followed the provided protocol within the EasySep™ Human CD14 Positive Selection Kit (Stemcell Technologies), which is designed to isolate CD14+ cells from peripheral blood mononuclear cells (PBMCs). Desired cells are hereby labelled with a bispecific tetrameric complex composed of monoclonal anti-CD14 and anti-dextran antibodies. By addition of dextran-coated magnetic particles, target cells get magnetically tagged and remain in the tube when this is placed into the purple EasySep™ magnet while unwanted cells are poured off. For the negative selection of monocytes, we used the EasySep™ Human Monocyte Enrichment Kit (also from Stemcell Technologies). Unwanted cells are labelled for removal with bispecific tetrameric antibody complexes recognizing the respective target antigen (CD2, CD3, CD16, CD19, CD20, CD56, CD66b, CD123, or glycophorin A) and dextran-coated magnetic particles. The antibody cocktail also contains an antibody to the human Fc receptor to prevent non-specific binding of monocytes. Labelled cells are separated using the purple EasySep™ magnet and remain in the tube while monocytes can be poured off into a new vial. The isolated neutrophils (purity >96–97%) and monocytes (purity >85%) were finally resuspended in PBS pH 7.4 containing 1 mg/mL of glucose and 1 mM CaCl2 (PGC buffer).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Cell Isolation Monocyte using EasySep™ Human Monocyte Enrichment Kit from STEMCELL technologies.

Manufacturer protocol

Download the product protocol from STEMCELL technologies for EasySep™ Human Monocyte Enrichment Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Cell Isolation Monocyte using EasySep™ Human Monocyte Enrichment Kit from STEMCELL technologies. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.