Human peripheral blood mononuclear cells (PBMCs) were isolated from human buffy coats from the New York Blood Center by density gradient centrifugation using Ficoll-Paque PLUS (GE Healthcare). Primary NK cells were extracted from PBMCs by negative immunomagnetic isolation using a NK cell isolation kit (Miltenyi Biotec) according to the manufacturer's protocol. Briefly, cells were incubated with NK Cell Biotin-Antibody Cocktail and NK Cell MicroBead Cocktail to label non-NK cells, followed by loading onto a LS Separation column (Miltenyi Biotec). The cell suspension was then placed in the magnetic field of a MACS Separator to separate labeled from non-labeled cells. The isolated human NK cells were subsequently cultured in Roswell Park Memorial Institute (RPMI) 1640 media supplemented with 10% fetal bovine serum, 1x penicillin/streptomycin and 100 U/mL IL-2 at 37°C in 5% CO 2 . The purity of the NK cell population was validated by flow cytometry using PE-Vio 770 anti-human CD56 (1:50; REA196 clone; Miltenyi Biotec) and FITC anti-human CD3 (1:50; REA613 clone; Miltenyi Biotec) antibodies.
Rea613 clone
Manufactured by Miltenyi Biotec
The REA613 clone is a laboratory equipment product offered by Miltenyi Biotec. It is designed to perform a core function, but a detailed description maintaining an unbiased and factual approach is not available.
Automatically generated - may contain errors
Lab products found in correlation
Nk cell biotin antibody cocktail, by Miltenyi Biotec (2 mentions)
Pe vio 770 anti human cd56, by Miltenyi Biotec (2 mentions)
Rea196 clone, by Miltenyi Biotec (2 mentions)
Fitc anti human cd3, by Miltenyi Biotec (2 mentions)
Nk cell microbead cocktail, by Miltenyi Biotec (2 mentions)
Ls separation column, by Miltenyi Biotec (2 mentions)
Ficoll paque plus, by GE Healthcare (2 mentions)
Nk cell isolation kit, by Miltenyi Biotec (2 mentions)
2 protocols using rea613 clone
1
Isolation and Culture of Human NK Cells
2
Isolation and Culture of Primary Human NK Cells
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Human peripheral blood mononuclear cells (PBMCs) were isolated from human buffy coats from the New York Blood Center by density gradient centrifugation using Ficoll-Paque PLUS (GE Healthcare). Primary NK cells were extracted from PBMCs by negative immunomagnetic isolation using a NK cell isolation kit (Miltenyi Biotec) according to the manufacturer’s protocol. Briefly, cells were incubated with NK Cell Biotin-Antibody Cocktail and NK Cell MicroBead Cocktail to label non-NK cells, followed by loading onto a LS Separation column (Miltenyi Biotec). The cell suspension was then placed in the magnetic field of a MACS Separator to separate labeled from non-labeled cells. The isolated human NK cells were subsequently cultured in Roswell Park Memorial Institute (RPMI) 1640 media supplemented with 10% fetal bovine serum, 1x penicillin/streptomycin and 100 U/mL IL-2 at 37°C in 5% CO2. The purity of the NK cell population was validated by flow cytometry using PE-Vio 770 anti-human CD56 (1:50; REA196 clone; Miltenyi Biotec) and FITC anti-human CD3 (1:50; REA613 clone; Miltenyi Biotec) antibodies.
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