VALIDATION IMAGES
Lane 1: mouse brain lysates Lane 2: mouse kidney lysates probed with Anti Smad3 Polyclonal Antibody, Unconjugated (bs-3484R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band 47kD. Observed band size: 47kD.
Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Ovary lysates; Lane 3: HT1080 cell lysates; Lane 4: Jurkat cell lysates probed with Smad2/3 Polyclonal Antibody, Unconjugated (bs-3484R) at 1:2000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
HUVEC cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad2/3 Polyclonal Antibody(bs-3484R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Heart lysates; Lane 3: Mouse Testis lysates; Lane 4: Mouse Skin lysates; Lane 5: Mouse Kidney lysates; Lane 6: Rat Cerebrum lysates; Lane 7: Rat Testis lysates; Lane 8: Rat Kidney lysates; Lane 9: Human HUVEC cell lysates; Lane 10: Human A549 cell lysates; Lane 11: Human Hela cell lysates; Lane 12: Human HT1080 cell lysates; Lane 13: Human A431 cell lysates probed with Smad2/3 Polyclonal Antibody, Unconjugated (bs-3484R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad2/3 Polyclonal Antibody(bs-3484R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Smad3) polyclonal Antibody, Unconjugated (bs-3484R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.