Mouse spleen cells were fixed with 4% PFA (10min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. The cells were then stained with SCF Polyclonal Antibody, Unconjugated (bs-0545R) at 1:33 for 30 min at room temperature. A Goat anti-rabbit IgG-AF647 secondary antibody was used at 1ug for 40 min at room temperature. Acquisition of 10,000 events was performed. Primary antibody staining (green) is compared to compared to unstained cells (purple), secondary only (light blue), and isotype control (orange).