15353-j-veluchamy

2 Harmonized NK FACS panels to study NK cell phenotype and function | 41 Blue 488nm PanKIR2D FITC NKVFS1 1:11 Miltenyi 130-098-689 NKG2A PE-Vio770 REA110 1:11 Miltenyi 130-105-647 NKG2C PE  REA205 1:11 Miltenyi 130-103-635 NKG2D PerCP-Cy5.5  1D11 1:11 Biolegend 320818 Red 633nm CD16 APC VEP13 1:11 Miltenyi 130-091-246 CD56 APC-Vio770  REA196 1:11 Miltenyi 130-100-694 Antibody-fluorochrome conjugates for NK phenotype panel were distributed across Violet, Blue and Red lasers. Back bone antibodies (grey shades) were assigned to Violet and Red lasers. VioGreen was used to gate on CD45+ cells and VioBlue in the Violet laser was used as a dump channel to exclude non-NK lymphocytes including dead cells. Red laser was used to gate on viable NK cells, followed by NK phenotype panel drop in markers (white shades) in the Blue laser. All antibodies except Sytox Blue (1:1000) were used at a dilution of 1:11. Table 3: Antibody specifications - NK function panel Laser Antibody Fluorochrome Clone Titration Manufacturer Catalogue No Violet 405nm CD45 VioGreen  5B1 1:11 Miltenyi 130-096-906 CD14 VioBlue  TÜK4 1:11 Miltenyi 130-094-364 CD19 VioBlue  LT19 1:11 Miltenyi 130-098-598 Sytox® Blue Dead cell marker 1:1000 Life technologies S11348 Blue 488nm CD25 VioBrightFITC 4E3 1:11 Miltenyi 130-104-274 CD107a PE  H4A3 1:11 Miltenyi 130-095-515 NKp44 PE-Vio770  2.29 1:11 Miltenyi 130-104-195 CD3 PerCP-Vio700  BW264/56 1:11 Miltenyi 130-097-582 TCRγδ PerCP-Vio700  11F2 1:11 Miltenyi 130-103-784 Red 633nm CD16 APC VEP13 1:11 Miltenyi 130-091-246 CD56 APC-Vio770  REA196 1:11 Miltenyi 130-100-694 Antibody-fluorochrome conjugates for NK function panel were distributed across Violet, Blue and Red lasers. Back bone antibodies (grey shades) were assigned to Violet and Red lasers. VioGreen was used to gate on CD45+ cells and VioBlue in the Violet laser was used as a dump channel to exclude CD14+, CD19+ and dead cells. Red laser was used to gate on viable NK cells. Blue laser was used to gate on CD3+, TCRγδ+ cells, non-conventional T cell subsets and NK function panel drop in markers (white shades). All antibodies except Sytox Blue (1:1000) were used at a dilution of 1:11. Laser Antibody Fluorochrome Clone Titration Manufacturer Catalogue No