VALIDATION IMAGES
Lane 1: HeLa; Lane 2: A431; Lane 3: K562 cell lysate probed with NFKB p65 (7G6) Monoclonal Antibody, unconjugated (bsm-33117M) at 1:300 overnight at 4°C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37°C.
Lane 1: Rela-KO cell lysates; Lane 2: Hela cell lysates probed with NFKB p65 (7G6) Monoclonal Antibody, Unconjugated (bsm-33117M) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: RELE-KO cell lysates; Lane 2: Hela cell lysates probed with NFKB p65 Monoclonal Antibody, Unconjugated (bsm-33117M) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Human MCF-7 cell lysates; Lane 2: Human MOLT-4 cell lysates; Lane 3: Human A549 cell lysates; Lane 4: Human Jurkat cell lysates; Lane 5: Mouse RAW264.7 cell lysates; Lane 6: Mouse NIH/3T3 cell lysates probed with NFKB p65 Monoclonal Antibody, Unconjugated (bsm-33117M) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Human HL-60 cell lysates probed with NFKB p65 Monoclonal Antibody, Unconjugated (bsm-33117M) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded Rat spleen; 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 NFKB p65 (7G6) Monoclonal Antibody, Unconjugated (bsm-33117M) at 1:200 overnight at 4°C, DAB staining.
Tissue/cell:MCF7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum,C-0005) at 37°C for 20 min; Antibody incubation with (NFKB p65) monoclonal Antibody, Unconjugated (bsm-33117M)1:100, 90 minutes at 37°C; followed by a CY3 conjugated Goat Anti-Mouse IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
25 ug total protein per lane of various lysates (see on figure) probed with NFKB p65 monoclonal antibody, unconjugated (bsm-33117M) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.