Immunofluorescence of HeLa cells labelling HIF-1-alpha with ab104263. Hela cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with ab104263 (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). HIF1 alpha immunoreactivity is localized to the cytoplasm and nucleus.
ab104263, at a 1/50 dilution, staining HIF-1-alpha in formalin fixed and paraffin embedded lung carcinoma by by Immunohistochemistry, followed by peroxidase conjugation of the secondary antibody and DAB staining.
Anti-HIF-1-alpha antibody (ab104263) at 1/50 dilution + CHO cell lysate at 35 µg