Paraformaldehyde-fixed, paraffin embedded (Human brain glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Cullin 1) Polyclonal Antibody, Unconjugated (bs-4208R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

4% Paraformaldehyde-fixed Hela(H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Cullin 1) polyclonal Antibody, unconjugated (bs-4208R) 1:50, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-0295G-FITC) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

The Hela (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. Primary Antibody (green): Rabbit Anti-Cullin 1 antibody (bs-4208R): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-0295G-FITC): 1 μg/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.