Nucleolin/C23 Rabbit Polyclonal Antibody

Blank control (blue): Hela (fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice). Primary Antibody: Rabbit Anti-Phospho-c-Fos (Ser32) /AF488 antibody, dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG/AF488 (orange), used under the same conditions.

Blank control: Molt4. Primary Antibody (green line): Rabbit Anti-Nucleolin/C23/FITC Conjugated antibody, dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG-FITC. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. The cells were stained with Primary Antibody for 30 min at room temperature. Acquisition of 20000 events was performed.

Tissue/Cell: rat uterus tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-Nucleolin/C23 Polyclonal Antibody, Unconjugated (orb2698) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.