VALIDATION IMAGES
Lane 1: rat brain lysates Lane 2: rat heart lysates probed with Anti Legumain Polyclonal Antibody, Unconjugated (bs-3907R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band 49kD. Observed band size: 49kD.
Moues kidney lysates probed with Legumain Polyclonal Antibody, unconjugated (bs-3907R) at 1:300 overnight at 4°C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37°C.
Mouse placenta lysates probed with Legumain Polyclonal Antibody, unconjugated (bs-3907R) at 1:300 overnight at 4°C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37°C.
Formalin-fixed and paraffin embedded human colorectal cancer labeled with Anti Legumain Polyclonal Antibody, Unconjugated (bs-3907R) followed by conjugation to the secondary antibody and DAB staining
HL-60 cells were fixed with 4% PFA for 10min at room temperature, 0.2% PBST for 20 min at room temperature. 5% BSA blocking buffer was used for 30 min at room temperature. Cells were then stained with Legumain Polyclonal Antibody (bs-3907R) at 1:33 dilution for 30 min at room temperature and then washed twice with 2% BSA in PBS. Secondary Antibody (PE-conjugated) incubation was performed for 40 min at room temperature. Acquisitions of 20,000 events were performed. The graph shows cells stained with primary antibody (green), and isotype control (orange).
Paraformaldehyde-fixed, paraffin embedded (mouse placenta); 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 (Legumain) Polyclonal Antibody, Unconjugated (bs-3907R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Lane 1: Human JAR cell lysates; Lane 2: Human MCF-7 cell lysates probed with Legumain Polyclonal Antibody, Unconjugated (bs-3907R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.