Anti nkp46 apc

Peripheral blood-derived and in vitro cultured mononuclear cell populations were identified by a combination of physical parameters and immunostaining with saturating concentrations of the following fluorochrome-conjugated mAbs: anti-CD3 PerCP (clone: SK7, cat #: 347344; BD Biosciences) or PerCP-Vio700 (clone: REA613, cat #: 130-109-465; Miltenyi Biotec), anti-CD56 APC (clone: B159, cat #: 555518; BD Biosciences) or APC-Vio770 (clone: REA 196, cat #: 130-100-694; Miltenyi Biotec), CD16 PE (clone: B73.1, cat #: 347617; BD Biosciences) or PE-Vio770 (clone: REA423, cat #: 130-106-706; Miltenyi Biotec), anti-FcεRIγ subunit FITC (polyclonal antibody, cat #: FCABS400F; Merck), anti-NKp46 APC (clone: REA808, cat #: 130-112-122; Miltenyi Biotec), anti-NKG2C PE (clone: 134591, cat #: FAB138P; R&D Systems), anti-PLZF PE (clone: Mags.21F7, cat #: 12-9320-82; ThermoFisher Scientific), anti-PD-1 PE (clone: EH12.2H7, cat #: 329906; BioLegend), and anti-PD-L1 APC (clone: 29E.2A3, cat #: 329708; BioLegend). Samples were stained for surface antigens for 30 min at 4°C, washed with PBS (Euroclone) containing 2% FCS and 2 mM EDTA (used for all washing steps), fixed with 2% paraformaldehyde for 20 min at room temperature (RT), washed, permeabilized with washing solution supplemented with 0.05% Triton-X 100 for 30 min at RT, and stained for intracellular antigens for 30 min at 4°C.