Immunofluorescence of MCF-7 cells labelling SMAD9 with ab124094. MCF-7 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with ab124094 (1:25, 1 h at 37℃). Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used as the secondary antibody (1:400, 50 min at 37℃). Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (7units/ml, 1 h at 37℃) and nuclei were counterstained with DAPI (blue) (10 µg/ml, 10 min). SMAD9 immunoreactivity is localized to the cytoplasm and nucleus.
Western blot - Anti-SMAD9 antibody (ab124094)
Anti-SMAD9 antibody (ab124094) at 1/100 dilution + A549 cell line lysates at 35 µg