Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.
Fluorescent confocal image of an SY5Y cell stained Phospho-STAT5a antibody. SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated Phospho-STAT5a primary antibody (1:200, 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). Nuclei were counterstained with Hoechst 33342 (blue) (10 ug/ml, 3 min)(not shown). Phospho-STAT5a immunoreactivity is localized exclusively to the nucleus.
Western blot of anti-Phospho-STAT5 Pabin HepG2 cell line lysate (35 ug/lane). Phospho-STAT5(arrow) was detected using the purified antibody.