Human neutrophils were isolated from the venous blood of healthy donors as described previously (Uriarte et al., 2011 ). In some experiments, further purification by negative magnetic selection was performed using the Easy Eights EasySep Magnet and human neutrophil enrichment kit (Stemcell Technologies, Vancouver, BC, Canada) (Miralda et al., 2020 (link); Vashishta et al., 2019 ).
Human neutrophil enrichment kit
Manufactured by STEMCELL
Sourced in Canada
The Human Neutrophil Enrichment Kit is a laboratory product designed to isolate and enrich human neutrophils from whole blood samples. It utilizes a density gradient centrifugation method to separate and collect the neutrophil fraction. The kit provides the necessary reagents and protocols to obtain a purified population of human neutrophils for further research and analysis.
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Lab products found in correlation
Easyeights easysep magnet, by STEMCELL (2 mentions)
Ficoll hypaque, by GE Healthcare (2 mentions)
Cd66abce microbead kit, by Miltenyi Biotec (1 mentions)
Human monocyte enrichment kit, by STEMCELL (1 mentions)
Ficoll paque premium 1, by GE Healthcare (1 mentions)
Human nk cell enrichment kit, by STEMCELL (1 mentions)
Hetasep solution, by STEMCELL (1 mentions)
6 protocols using human neutrophil enrichment kit
1
Isolation of Human Neutrophils
2
Isolation of Human Neutrophils
3
Isolation of Human Neutrophils
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Neutrophils were isolated by means of negative selection magnetic isolation using the Human Neutrophil Enrichment Kit (StemCell Technologies) according to the manufacturer's guidelines. Briefly, total white cells were incubated with EasySep Human Neutrophil Enrichment cocktail for 10 minutes at 4°C. EasySep nanoparticles were then added to the mixture and were incubated for 10 minutes at 4°C. The suspension was then mixed and brought to a total volume of 2.5 mL. The tube was then inserted into the EasySep magnet, and with one continuous motion, the tube was inverted, and the contents were poured into a fresh 5 mL polystyrene tube. Cell count and viability were determined using the trypan blue exclusion method.
4
Isolation and Characterization of Neutrophils and LDGs from P. vivax Patients
5
Isolation of Human Blood Immune Cells
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Human peripheral blood monocytes, neutrophils and NK cells were isolated from healthy male adult donor aphaeresis cones (National University Hospital, Blood Donation Centre, Singapore). This work was approved by the Institutional Review Board (IRB), NUS-IRB B-14-063E, National University of Singapore (NUS). Monocytes and NK cells were isolated as described previously [59 (link)]. Briefly, the buffy coat, which contained citrate-phosphate-dextrose (CPD) as anticoagulant, was diluted four times with PBS containing 2% FBS and 1 mM EDTA, and the mononuclear fraction was obtained via density gradient centrifugation with Ficoll-Paque Premium 1.073 (GE Healthcare). From the mononuclear fraction, the monocyte and NK cell populations were enriched with the Human Monocyte Enrichment Kit and Human NK Cell Enrichment Kit (Stemcell), respectively. For neutrophil isolations, buffy coats were diluted using Hank’s Balanced Salt Solution (HBSS; Life Technologies). Red blood cells were sedimented by gravity using Hetasep solution (Stemcell). Leukocytes were harvested and neutrophils were isolated using Human Neutrophil Enrichment kit (Stemcell).
6
Isolation of Neutrophils from Infected Patients
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Neutrophils from acutely and chronically infected patients and from HDs were isolated with a Ficoll-Hypaque (GE Biosciences) gradient followed by a human neutrophil enrichment kit (STEMCELL Technologies) according to the manufacturer’s instructions. Neutrophil purity was assessed by flow cytometry.
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