Tissue/cell: human glioma tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum) at 37℃ for 20 min; Incubation: Anti-PDCD4 Polyclonal Antibody, Unconjugated(PC1608) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(A10026) and DAB staining

Tissue/cell: mouse kidney tissue;4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue) was used to stain the cell nuclei

Sample: Lane1: Kidney at 1:3000 dilution; Predicted band size : 50kD Observed band size : 50kD

HepG2 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 (PDCD4) polyclonal Antibody, Unconjugated (PC1608) 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.

Blank control（black line）:HepG2. Primary Antibody (green line): Rabbit Anti-PDCD4 antibody (PC1608) Dilution:2ug/Test; Secondary Antibody（white blue line）: Goat anti-rabbit IgG-FITC Dilution: 0.5ug/Test. Isotype control（orange line）: Normal Rabbit IgG 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.