Table 2. Antibody specifications - NK phenotype panel.
| Laser | Antibody | Fluorochrome | Clone | Titration | Manufacturer | Catalogue No |
|---|---|---|---|---|---|---|
| Violet 405 nm | CD45 | VioGreen | 5B1 | 1:11 | Miltenyi | 130-096-906 |
| CD3 | VioBlue | BW264/56 | 1:11 | Miltenyi | 130-094-363 | |
| TCRγδ | VioBlue | 11F2 | 1:11 | Miltenyi | 130-101-557 | |
| 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 488 nm | 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 633 nm | 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. Backbone antibodies (Bold) 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.