Pe cyanine7 anti human cd16 antibody

Four PBMC vials from patients with lupus were taken from our UNCOVR database, and four healthy donors, from Bloodworks and Stemcell, were taken as well. These samples were assayed with the following antibodies: APC anti-human CD57 Antibody (BioLegend #359609), FITC anti-human CD56 (NCAM) Antibody (BioLegend #318303), BD OptiBuild BUV395 anti-human CD8 (BD Biosciences #740303), Brilliant Violet 421 anti-human CD159a (NKG2A) antibody (BioLegend #375139), PE anti-human CD159c (NKG2C) Antibody (BioLegend #375003), APC/Cyanine7 anti-human CD127 (IL-7Rα) Antibody (BioLegend #351347), Brilliant Violet 785 anti-mouse/human KLRG1 (MAFA) antibody (BioLegend #138429), PE/Cyanine7 anti-human CD16 antibody (BioLegend #302015), and LIVE/DEAD Fixable Aqua Dead Cell Stain Kit, for 405 nm excitation (ThermoFisher #L34965). PBMC samples were resuspend from −80°C in R10 media to remove and dilute freezing media. Post-washing each pellet was resuspended and washed in one mL ice-cold PBS then resuspended in live-dead stain for 30 min at 4°C. Stained cells were then washed in one mL ice-cold PBS and stained with the surface-protein antibody master mix for 30 min at 4°C. Stained cells were then washed with cell staining buffer, the same that was utilized for the master mix, and resuspended in 100 μL and run on BD FACSymphony A5 Cell Analyzer at Fred Hutchinson Cancer Center.

Four PBMC vials from patients with lupus were taken from our UNCOVR database, and four healthy donors, primarily from Bloodworks, were taken as well. These samples were assayed with the following antibodies: APC anti-human CD57 Antibody (BioLegend #359609), FITC anti-human CD56 (NCAM) Antibody (BioLegend #318303), BD OptiBuild^™ BUV395 anti-human CD8 (BD Biosciences #740303), Brilliant Violet 421^™ anti-human CD159a (NKG2A) antibody (BioLegend #375139), PE anti-human CD159c (NKG2C) Antibody (BioLegend #375003), APC/Cyanine7 anti-human CD127 (IL-7Rα) Antibody (BioLegend #351347), Brilliant Violet 785^™ anti-mouse/human KLRG1 (MAFA) antibody (BioLegend #138429), PE/Cyanine7 anti-human CD16 antibody (BioLegend #302015), and LIVE/DEAD^™ Fixable Aqua Dead Cell Stain Kit, for 405 nm excitation (ThermoFisher #L34965). PBMC samples were resuspend from − 80°C in R10 media to remove and dilute freezing media. Post-washing each pellet was resuspended and washed in one mL ice cold PBS then resuspended in live-dead stain for 30 minutes at 4°C. Stained cells were then washed in one mL ice-cold PBS and stained with the surface-protein antibody master mix for 30 minutes at 4°C. Stained cells were then washed with cell staining buffer, the same that was utilized for the master mix, and resuspended in 100 μL and run on BD FACSymphony^™ A5 Cell Analyzer at Fred Hutchinson Cancer Center.