VALIDATION IMAGES
Lane 1: HeLa lysates; Lane 2: SHG-44 lysates; Lane 3: SW480 Lysates; probed with Stat3 (6G5) Monoclonal Antibody (bsm-52235R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.
CRC cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS and stained with Stat3 (6G5) Monoclonal Antibody (bsm-52235R) 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 pancreas; 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 Stat3 (6G5) Monoclonal Antibody (bsm-52235R) at 1:50 overnight at 4°C, followed by a conjugated secondary and DAB staining.
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 Stat3 Antibody(bsm-52235R)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: Hela cell lysates; Lane 2: A431 cell lysates probed with Stat3 Monoclonal Antibody, Unconjugated (bsm-52235R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.