VALIDATION IMAGES
MCF7 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 FOXP3 (12B1) Monoclonal Antibody(bsm-52079R)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).
Paraformaldehyde-fixed and paraffin-embedded Human tonsil tissue incubated with FOXP3 (12B1) Monoclonal Antibody (bsm-52079R) at 1:200, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
Paraformaldehyde-fixed, paraffin embedded Human tonsil; 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 FOXP3 (12B1) Monoclonal Antibody, Unconjugated (bsm-52079R) at 1:200 overnight at 4°C, DAB staining.
Lane 1: 293T cell lysates probed with FOXP3 Monoclonal Antibody, Unconjugated (bsm-52079R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
MCF-7 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 FOXP3 (12B1) Monoclonal Antibody(bsm-52079R)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).