Fluorescent confocal image of HeLa cells stained KLF4 (S457) antibody. HeLa cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated KLF4 (S457) primary antibody (1:100, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Cytoplasmic actin was counterstained with Alexa Fluor 555 (red) conjugated Phalloidin (5.25 mu M, 25 min). Note the highly specific localization of the phosphorylated KLF4 immunoreactivity to the nuclei but not the cytoplasm.
Fluorescent confocal image of SY5Y cells stained KLF4 (S457) antibody. SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated KLF4 (S457) primary antibody (1:100, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Cytoplasmic actin was counterstained with Alexa Fluor 555 (red) conjugated Phalloidin (5.25 mu M, 25 min). Note the highly specific localization of the phosphorylated KLF4 immunoreactivity to the nuclei.
Immunofluorescent of U251 cells, using KLF4 Antibody. Antibody was diluted at 1:25 dilution. Alexa Fluor 488-conjugated goat anti-rabbit lgG at 1:400 dilution was used as the secondary antibody (green). Cytoplasmic actin was counterstained with Dylight Fluor 554 (red) conjugated Phalloidin (red).
Western blot of KLF4-S457 in MDA-MB231, NCI-H460 cell line lysates (35 ug/lane). KLF4 (arrow) was detected using the purified antibody.