Anti human igg fc pe

Efficiency of retroviral transduction was determined by flow cytometry. All staining and washing was performed with PBS + 1% FBS. For the IL13Rα2-CAR and CD19-CAR, T cells were washed, incubated with 7 μl of recombinant IL13Rα2-F_c fusion protein (R&D Systems) or 3 μl of recombinant human CD19-F_c fusion protein (R&D Systems) at room temperature for 1 hour, washed, and stained with 1 μl of anti-human IgG F_c PE (SouthernBiotech) for 30 min at 4°C. All other transgene staining was performed at 4°C for 30 min with the following antibodies: HER2-CAR: anti-mouse Fab AF647 (Jackson ImmunoResearch; 2 μl per test); EphA2-CAR: anti-human CD19 PE (Beckman Coulter; 3 μl per test) or anti-human Fab AF647 (Jackson ImmunoResearch; 2 μl per test); and IL-15 vector: anti-human nerve growth factor receptor (NGFR) BV421 (BD Biosciences; 5 μl per test). Samples were acquired on a BD FACSLyric or BD FACSCanto, and list mode files were analyzed using FlowJo version 10.5.3 (BD Biosciences).