VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded rat kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37°C for 20min; Antibody incubation with AGEs Polyclonal Antibody, Unconjugated (bs-1158R) at 1:200 overnight at 4°C, followed by a conjugated secondary and DAB staining.
MCF7 cells were incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with AGEs Antibody(bs-1158R)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).
Lane 1: Mouse Kidney tissue lysates; Lane 2: Mouse Lung tissue lysates; Lane 3: Mouse Large intestine tissue lysates; Lane 4: Rat Testis tissue lysates ; Lane 5:Rat Lung tissue lysates ; Lane 6: Rat Large intestine tissue lysates; Lane 7: Human U87MG cell lysates probed with AGEs Polyclonal Antibody, Unconjugated (bs-1158R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
25 ug total protein per lane of various lysates (see on figure) probed with AGEs polyclonal antibody, unconjugated (bs-1158R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
25 ug total protein per lane of various lysates (see on figure) probed with AGEs polyclonal antibody, unconjugated (bs-1158R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.