Pe goat anti mouse igg antibody

After 96 h of infection with CAR lentiviral, the T cells were analyzed for CAR expression using flow cytometry and immunofluorescence assay. The UCAR-T cells were blocked with human Fc blocker (cat# 564220, BD), followed by consecutive incubation with 5 µg/mL BCMA-mFc (CAR-hu388, CAR-h31 and CAR-h32) and CD47-hFc (CAR-hu404, CAR-h31 and CAR-h32) proteins, APC anti-human IgG Fc antibody (cat# 410711, Biolegend) and PE-goat anti-mouse IgG antibody (cat# 405307, Biolegend). PE-goat IgG isotype control antibody (cat# 403004, Biolegend) and APC Rat IgG2a, κ isotype control antibody (cat# 400511, Biolegend) was used as negative controls.

To analyze the populations of endothelial cells and cardiomyocytes in a given cell sheet, flow cytometry was performed using a Gallios cytometer (Beckman Coulter, CA, USA). The suspension containing the isolated cells was incubated with a mouse monoclonal anti-rat CD31 antibody (MCA1334G; AbD Serotec) at a dilution of 1/100 for 30 min at room temperature; cardiomyocytes were incubated with an anti-troponin T antibody (Cardiac Isoform Ab-1, Clone: 13-11; Thermo Scientific, Waltham, MA, USA) at a dilution of 1/100 for 30 min at room temperature. After being washed with PBS, cells were incubated with PE Goat anti-mouse IgG antibody (405307; BioLegend, CA, USA) at a dilution of 1/100 for 30 min at room temperature. After the antibody reaction, the cells that reacted were analyzed.