Phospho-Cyclin B1 (Ser133) Rab

Blank control (blue line): A549 (blue). Primary Antibody (green line): Rabbit Anti-Phospho-Cyclin B1 (Ser133) antibody (orb5927), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): F (ab')2 fragment goat anti-rabbit IgG-FITC, dilution: 1 µg/Test. Protocol, The cells were fixed with 2% Fixative (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Hela cell, 4% Fixative-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Phospho-Cyclin B1 (Ser133)) polyclonal Antibody, Unconjugated (orb5927) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.