VALIDATION IMAGES
Formalin-fixed and paraffin embedded mouse embryo tissue labeled with Anti- IKK beta Polyclonal Antibody, Unconjugated (bs-4880R) at 1:200 followed by conjugation to the secondary antibody
A431 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 IKK_/_ Polyclonal Antibody(bs-4880R)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, paraffin embedded Rat bladder; 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 IKK beta Polyclonal Antibody, Unconjugated (bs-4880R) at 1:200 overnight at 4°C, DAB staining.
25 ug total protein per lane of various lysates (see on figure) probed with IKK beta polyclonal antibody, unconjugated (bs-4880R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with IKK beta Polyclonal Antibody, Unconjugated (bs-4880R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with IKK beta Polyclonal Antibody, Unconjugated (bs-4880R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.