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; Incubation: Anti-PCGF4/BMI1 Antibody, conjugated (SL2999R-FITC) 1:400, 1.5 hours at 37°C; DAPI (5ug/ml, blue, SLC0033) was used to stain the cell nuclei.
Blank control (Black line): HUVEC (Black).
Primary Antibody (green line): Rabbit Anti-PCGF4 BMI1 antibody (SL2999R-FITC)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG-FITC .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells then were 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. Acquisition of 20,000 events was performed.
Blank control (blue line): Mouse kidney (Black).
Primary Antibody (green line): Rabbit Anti-PCGF4 BMI1/FITC antibody (SL2999R-FITC)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG-FITC .
Protocol
The cells were fixed with 4% PFA (5 min at -20℃) and then permeabilized with ice-cold mehtanol for 20 min at -20℃.Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions followed by the antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.
