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Blank control:Raw264.7. Isotype Control: Rabbit IgG-FITC(bs-0295P-FITC) Dilution: 1ug/Test; Primary Antibody (green line): Rabbit Anti- Fas Ligand antibody (bs- 0216R-FITC) Dilution: 1ug/Test; Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Isotype control antibody (orange line) was rabbit IgG .Acquisition of 20,000 events was performed.