Western blot - Anti-NF-kB p65 antibody [E379] (ab32536) Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: NF-kB p65 knockout HAP1 cell lysate (20 µg) Lane 3: HeLa cell lysate (20 µg) Lane 4: A431 cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab32536 observed at 70 kDa. Red - ab8245 loading control, observed at 37 kDa.

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NF-kB p65 with purified ab32536 at 1:100 dilution. Cells were fixed in 100% methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling NF-kB p65 with ab32536 at 1/5000 (0.098 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse spleen. The section was incubated with ab32536 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

Western blot - Anti-NF-kB p65 antibody [E379] (ab32536) All lanes : Anti-NF-kB p65 antibody [E379] (ab32536) at 1/1000 dilution Lane 1 : Human fetal brain lysates Lane 2 : Human fetal kidney lysates Lane 3 : Human fetal lung lysates Lysates/proteins at 20 µg per lane. Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution Predicted band size: 65 kDa Observed band size: 65 kDa Blocking/Diluting buffer and concentration: 5% NFDM/TBST Exposure time: 37 second for Lane1 and 3 seconds for Lanes 2 and 3 Normal brain might express low level of p65 (PMID: 21479220)