Sample: Lane 1: HepG2 cell lysates Lane 2: Human testis tissue lysates Lane 3: A549 cell lysates Lane 4: A431 cell lysates Lane 5: Jurkat cell lysates Lane 6: Mouse brain tissue lysates Primary: Anti-EEF1E1 (bsm-51605M) at 1/2000~4000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 21 kD Observed band size: 18 kD

Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (EEF1E1) monoclonal Antibody, Unconjugated (bsm-51605M) 1:25, 90 minutes at 37°C; followed by a conjugated Goat Anti-Mouse IgG antibody at 37°C for 90 minutes, Dylight® 554 Phalloidin (red) was used to stain the cell Cytoplasmic actin.The nuclear counter stain is DAPI (blue).

Blank control: Hela. Primary Antibody (green line): Mouse Anti-EEF1E1 antibody (bsm-51605M) Dilution: 1:25; Isotype Control Antibody (blue line): Mouse IgG Secondary Antibody : Goat anti-mouse IgG-AF488 Dilution: 1:400 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. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.