Immunohistochemistry of Mouse anti-AKT pS473 antibody. Tissue: human prostate tissue. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: AKT pS473 antibody at 20 µg/mL for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: AKT pS473 is nuclear and occasionally cytoplasmic. Staining: AKT pS473 as precipitated red signal with hematoxylin purple nuclear counterstain.
Immunohistochemistry using Anti-AKT pS473 monoclonal antibody shows detection of phosphory-lated AKT pS473 in human prostate tissue. The antibody was used at 20 ug/mL. The staining is much stronger than the weak basal level of phosphorylation in normal prostate tissue. Tissue was formalin fixed and paraffin embedded. No pre-treatment of sample was required. Signal was developed using Dako's Techmate streptavidin-biotin reagents. Personal communication, Glenna Burmer, Lifespan Biosciences, Seattle, WA.