Immunofluorescence microscopy: Rabbit anti-AKT was used at a 1:80 dilution to stain cultured neonatal rat cardiomyocytes that express a nuclear-targeted AKT construct. Anti-AKT staining appears green. Actin filaments are labeled red using a Texas-red™ conjugated phalloidin.
Immunoblotting: Rabbit anti-AKT was used at a 1:500 dilution to detect AKT by Western blot. Approximately 20 ug of an NIH/3T3 whole cell lysate was loaded on a 10% NuPage SDS-PAGE gel using MOPS buffer. After washing, a 1:10,000 dilution of HRP conjugated Gt-a-Rabbit IgG (R1454HRP) preceded color development using Pierce Chemical's SuperSignal™ substrate.
Immunohistochemistry. Rabbit anti-AKT was used at a 1:1,000 dilution to detect AKT by immunohistochemistry..,.In panel (A) normal colon tissue is stained. Panel (B) shows tumor tissue. Tissue was formalin-fixed and paraffin embedded.