Sample:
Lane 1: Mouse Prostate tissue lysates
Lane 2: Mouse Placenta tissue lysates
Lane 3: Rat Prostate tissue lysates
Lane 4: Rat Placenta tissue lysates
Lane 5: Human HepG2 cell lysates
Lane 6: Human MCF-7 cell lysates
Lane 7: Human SH-SY5Y cell lysates
Primary: Anti-TGF beta 3 propeptide (bs-0099R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 30/47 kDa
Observed band size: 47 kDa

Tissue/cell: pig bile duct; 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,C-0005) at 37℃ for 20 min;
Incubation: Anti-TGF-beta-3 Polyclonal Antibody, Unconjugated(bs-0099R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control: SHSY5Y.
Primary Antibody (green line): Rabbit Anti-TGF beta 3 propeptide antibody (bs-0099R)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
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.