Anti cd19 apc

PBMC staining and analysis were as described [3 (link)]. Unless otherwise specified, mAbs were from BD Pharmingen and included anti-CD3-Alexa Fluor 700 (UC-HT1), anti-CD4-APC (RPA-T4), anti-CD8-PacificBlue (RPA-T8), anti-CD11c-FITC (BU15, Thermo Scientific), anti-CD14-APC (RMO52, Beckman Coulter), anti-CD16-PacificBlue (3G8), anti-CD19-APC (HIB19) and anti-CD20-FITC (L27), anti-CD56-APC (N901, Beckman Coulter), and anti-NKG2D-PE (1D11; [2 (link)]). Anti-NKG2DL mAb were anti-MICA/B-PE (6D4), anti-ULBP1-PE (170818, R & D Systems), anti-ULBP2/5/6-PE (165903; R & D Systems), anti-ULBP3-PE (166510; R & D Systems), and anti-ULBP4-PE (1H1; [12 (link)]). In some cases, viable cell numbers were limiting resulting in partial data sets.

Cell suspension (1–2 × 10⁶) was incubated with antibodies for 20 min at 4 °C in 100 µL of phosphate buffered saline (PBS). The following anti-human monoclonal antibodies, all fluorescein isothiocyanate (FITC)- or phycoerythrin (PE)- or allophycocyanin (APC)-conjugated, were used at 1:10 dilution: anti-CD90 FITC (ref: IM1839U), anti-CD73 PE (ref: B68176), anti-CD105 PC7 (ref: B43293), anti-CD45 FITC (ref: IM0647), anti-CD34 FITC (ref: IM1870), anti-CD14 FITC (ref: B36297), anti-HLA-DR PE (ref: IM1639), anti-CD19 APC (ref: IM2470), anti-CD31 FITC (ref: IM1431U) (Beckman Coulter, Brea, CA, USA), anti-CD146 APC (clone: REA773), anti-SUSD2 APC (clone: W5C5), and anti-EPCAM FITC (clone: HEA-125) (Miltenyi Biotech, Bergisch Gladbach, Germany). As negative control, cells were incubated without antibodies. Labelled cells were washed with PBS and analyzed using Navios cytometer (Beckman Coulter, CA, USA).