HepG2 cells were fixed with 4% PFA for 10min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with BDNF Polyclonal Antibody(bs-4989R) at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA  in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed.  Cells stained with primary antibody (green), no antibody control control (blue), secondary antibody only (light blue) and isotype control (orange).