VALIDATION IMAGES
Formalin-fixed and paraffin embedded rat brain labeled with Rabbit Anti pro-caspase-3 Polyclonal Antibody, Unconjugated (bs-2593R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
L1 mouse brain lysates L2 mouse liver lysates probed with Anti pro-caspase-3 Polyclonal Antibody, Unconjugated (bs-2593R) at 1:200 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 for 90 min at RT. Predicted band 30kD. Observed band size:30kD.\n
Hela cells were fixed with 70% ice-cold methanol overnight at 4℃,0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Caspase 3 Polyclonal Antibody 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 incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed.
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 Caspase 3 Polyclonal Antibody, Unconjugated (bs-2593R) at 1:400 overnight at 4°C, DAB staining.
HL-60 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Caspase 3 Polyclonal Antibody(bs-2593R)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 Spleen lysates; Lane 2: Mouse Lung lysates probed with Caspase 3 Polyclonal Antibody, Unconjugated (bs-2593R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.