Anti cd56 phycoerythrin

A single cell suspension of primary NK cells was prepared in Cell Staining Buffer (cat. no. 420201; BioLegend, Inc.). Cells were pre-incubated with 5 µl of Human TruStain FcX™ (cat. no. 422301; BioLegend, Inc.) per 100 µl (1×10⁶ cells) of cell suspension for 5–10 min at room temperature to block Fc-receptors. The samples were centrifuged at 350 × g for 5 min at room temperature and the supernatant was discarded. Cells were incubated with the following fluorescently labelled antibodies: Anti-CD3-FITC (5 µl/1×10⁶ cells; cat. no. 300305; BioLegend, Inc.) or anti-CD56-phycoerythrin (5 µl/1×10⁶ cells; cat. no. 362507; BioLegend, Inc.) and incubated on ice for 15–20 min in the dark. Cells were washed two times with 2 ml of Cell Staining Buffer and centrifuged at 350 × g for 5 min at room temperature. Cells were analyzed by flow cytometry using a BD FACSCalibur (Becton-Dickinson and Company) and data was analyzed using FCS Express 6.06.0022 (De Novo Software).