VALIDATION IMAGES
Western blot analysis of Phospho-Smad3(S423/S425) on different lysates using anti-Phospho-Smad3(S423/S425) antibody at 1:1,000 dilution. Positive control: Lane 1: A431 cell lysates-treated with TGF β Lane 2: A431 cell lysates-untreated
A431 were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS and stained with Smad3(S423/S425) (5C5) Monoclonal Antibody (bsm-52205R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.
Paraformaldehyde-fixed, paraffin embedded mouse skin section; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37°C for 20min; Antibody incubation with Smad3(S423/S425) (5C5 ) Monoclonal Antibody (bsm-52205R) at 1:50 overnight at 4°C, followed by a conjugated secondary and DAB staining.
Lane 1: A549 cell lysates probed with Smad3(S423/S425) (5C5) Monoclonal Antibody, Unconjugated (bsm-52205R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Siha cell lysates probed with Smad3(S423/S425) (5C5) Monoclonal Antibody, Unconjugated (bsm-52205R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Hela cell lysates probed with Smad3(S423/S425) (5C5) Monoclonal Antibody, Unconjugated (bsm-52205R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Mouse Lung lysates; Lane 2: A549 cell lysates; Lane 3: Hela cell lysates probed with Phospho-Smad3 (Ser423 + Ser425) Monoclonal Antibody, Unconjugated (bsm-52205R ) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.