VALIDATION IMAGES
Lane 1: Hela lysates; Lane 2:A431 lysates; Lane 3: PC12 lysates probed with Smad2 (9A3) Monoclonal Antibody (bsm-52223R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.
Hela cells were stained with Smad2 (9A3) Monoclonal Antibody (bsm-52223R) at [1:200] incubated for overnight at 4C followed by secondary antibody incubation, DAPI staining of the nuclei and detection.
A431 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 (9A3) Monoclonal Antibody(bsm-52223R)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: MouseTestis lysates; Lane 2: Human A431 cell lysates; Lane 3: Human HepG2 cell lysates; Lane 4: Human HL60 cell lysates; Lane 5: Human Jurkat cell lysates probed with Smad2 monoclonal Antibody, Unconjugated (bsm-52223R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded Rat kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4°C, DAB staining.
A549 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 (Smad2) monoclonal Antibody, Unconjugated (bs-52223R) 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.