A488 donkey anti mouse igg

Pipette blood samples into 1.5 mL EP tubes. Spin at 4°C for 10 minutes at 300g, then discard the supernatant. Add 200 μL of PBS/0.5% BSA and vortex. Next, incubate for 10 minutes with primary antibodies at 4°C. Spin at 4°C for 10 minutes at 300g, discard the supernatant, and add 200 μL of PBS/0.5% BSA. Incubate for 10 minutes with fluorescent-labeled secondary antibodies and DAPI at 4°C. Spin at 4°C for 10 minutes at 300g, discard the supernatant, and smear. Seal the slices with an anti-fluorescence quencher. Immunofluorescent images were captured using the fluorescence microscope (Leica DMI300B). The fluorescent intensity was acquired using the software ImageJ. The primary antibodies used in this study include: Rat-anti-human CD19 (Invitrogen, #11-0193-82), Mouse-anti-human CD27 (Sigma-Aldrich, #SAB4700131), Rabbit-anti-human MIF (Affinity, #DF6404), Mouse-anti-human CD68 (Invitrogen, #14-0688-82), Rat-anti-human CD44 (Invitrogen, #14-0441-82), and Goat-anti-human CD74 (Invitrogen, #PA5-143198). The secondary antibodies used in this study include: A594 Goat Anti-Rabbit IgG (Affinity, #S0006), A647 Goat Anti-Mouse IgG (Bi Yuntian, #A0473), A488 Donkey-anti-Mouse IgG (Invitrogen, #A21202), A647 Donkey anti-Rabbit IgG (Invitrogen, #A32795), and DAPI (Invitrogen, #D1306).