Peripheral blood was collected in sodium heparin tubes and processed within 4 hours of blood draw. HetaSep™ (Stem Cell Technologies; Vancouver, BC) was used to deplete red blood cells. To isolate neutrophils and PBMCs, total leukocytes were placed over Histopaque 1077 (Sigma; St. Louis, MO) and centrifuged (400g, 30 minutes, no brake). Using magnetic microbeads, CD14+ monocytes were positively selected from PBMCs (EasySep™ Stem Cell Technologies), and enriched leukocytes were isolated from the CD14- PBMCs with a human CD4 T cell enrichment cocktail (Stem Cell Technologies).46 (link), 47 (link) Neutrophils were isolated from red blood cell/granulocyte pellet after Histopaque separation and red cell lysis, and lysed for RNA or resuspended in Hank’s Buffered Saline (HBSS, Gibco; Gaithersburg, MD) for in vitro activation experiments. Low-density neutrophils (LDNs) were enriched from PBMCs using the human neutrophil isolation kit (Stem Cell Technologies).
Human neutrophil isolation kit
Manufactured by STEMCELL
Sourced in United States
The Human Neutrophil Isolation Kit is a laboratory product that allows for the isolation and purification of human neutrophils from whole blood samples. It utilizes density gradient centrifugation to separate and enrich the neutrophil cell population.
Automatically generated - may contain errors
Lab products found in correlation
Hetasep, by STEMCELL (1 mentions)
Human cd4 t cell enrichment cocktail, by STEMCELL (1 mentions)
Histopaque, by Thermo Fisher Scientific (1 mentions)
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions)
Histopaque 1077, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
2 protocols using human neutrophil isolation kit
1
Isolation of Neutrophils and PBMCs
2
Leukemia Cell Line Culture and Treatment
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The human APL cell lines (HL-60 and NB4), T-ALL cell lines (Jurkat and CEM-C7), and monocytic leukemia cell line (THP-1) were purchased from Jennio Biotechnology Co., Ltd (Guangzhou, Guangdong, CHN). Blood samples were obtained from healthy volunteer. Neutrophils were isolated with human neutrophil isolation kit (STEMCELL, CA, USA). PBMCs and monocytes were extracted with isolation kit (Solarbio, Beijing, China). Cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (Gibco, Waltham, MA, USA) at 37 °C in a humidified incubator with 5% CO2. Cells were cultured in culture medium (normal control), and were treated with 2F5 or IgG (negative control) with different concentrations (20, 40, and 80 μM) for 4, 8, 12, and 16 days. Every 4 days, the cultures were established by centrifugation and then the cell pellets were resuspended in fresh corresponding medium respectively.
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