Immunofluorescent analysis of U251 cells, using KLF4 Antibody. BD-PB2104 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 analysis of KLF4-S457 in MDA-MB231, NCI-H460 cell line lysates (35ug/lane). KLF4 (arrow) was detected using the purified Pab.

Fluorescent confocal image of HeLa cells stained with BD-PB2104 KLF4 (S457) antibody. HeLa cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated with BD-PB2104 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 μM, 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 with BD-PB2104 KLF4 (S457) antibody. SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated with BD-PB2104 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 μM, 25 min). Note the highly specific localization of the phosphorylated KLF4 immunoreactivity to the nuclei.