Anti human cd19 apc

Mice were bled from the submandibular vein, the erythrocytes in the blood were lysed with ACK lysis buffer, and the sample stained with the following antibodies: anti-mouse-CD45-PECy7 (BioLegend, Munich, Germany; clone 30-F11), anti-human-CD45-Pacific Orange (Thermo Fisher, Darmstadt, Germany; clone HI30), anti-human-CD19-APC (BD Pharmigen, Heidelberg, Germany; clone HIB19), anti-human-CD3-Pacific Blue (BD Pharmigen, Heidelberg, Germany; clone UCHT1), anti-human-CD4-PE (BD Pharmigen, Heidelberg, Germany; clone L120), anti-human-CD8-FITC (BD Pharmigen, Heidelberg, Germany; clone SK1), anti-human-CD16-AF700 (BD Pharmigen, Heidelberg, Germany; clone CD2F1). The samples were washed and analyzed in FACS Buffer (PBS containing 2% FBS and 2 mM EDTA pH = 8.0). Mouse tissue lymphocytes were isolated as detailed above and 1 × 10⁶ cells were used for FACS staining. FACS data were acquired using a BD FACSAria Fusion flow cytometer and sample analysis was done on the FlowJo v10.2 software with final graphs and statistical analysis being performed in GraphPad Prism 7.

DFBL-18689-V2 was acquired from the Public Repository of Xenografts (PRoXe). PDX cells (0.25–1 × 10⁶) were injected IV into 6–8-week-old NSG mice, and mice were sacrificed on day 26–31 after injection. Single-cell suspensions were obtained from spleens, and erythrocytes were lysed (ACK Lysing Buffer). Cells were stained with combinations of the following antibodies: anti-mouse CD45-APC (Cat#559864, clone 30-F11), anti-mouse CD45.1-V450 (Cat#560520, clone A20), anti-mouse CD45.2-V450 (Cat#560697, clone 104) anti-human IgM-APC (Cat#551062, clone G20-127), anti-human CD20-FITC (Cat#555622, clone 2H7), and anti-human CD19-APC (Cat#555415, clone HIB19) (all from BD Pharmigen). For proliferation analysis, cells were fixed and permeabilized using the Intracellular Fixation and Permeabilization Buffer Set (eBiosience) and stained with anti-ki67 (Abcam; Cat#ab16667); staining was revealed with alexa fluor 647 goat anti-rabbit IgG (H + L) (L.T.) (ThermoFisher, A-21245). Live cells were detected with DAPI (Sigma-Aldrich) or with LIVE/DEAD Fixable Yellow Dead Cell Stain (Thermo Fisher, L34959). Labeled cells were acquired with a LSRFortessa High-Parameter Flow Cytometer and analyzed with FlowJo V10.4.2 software.