Immunohistochemistry analysis of FFPE human breast carcinoma tissue section using A1125-04C (1:250) after 20 minutes of heat induced antigen retrieval (HIER) with 10 mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block on a Bond Rx autostainer (Leica Biosystems). The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This antibody generated a diffused cytoplasmic staining of phosphor-AKT (Ser-473) in the cancer cells as well as the stromal cells. Some cancer cells depicted nuclear stianing also. Staining was performed by Histowiz.

Immunohistochemistry analysis of FFPE human breast carcinoma tissue section using A1125-04C (1:250) after 20 minutes of heat induced antigen retrieval (HIER) with 10 mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block on a Bond Rx autostainer (Leica Biosystems). The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This antibody generated a diffused cytoplasmic staining of phosphor-AKT (Ser-473) in the cancer cells as well as the stromal cells. Staining was performed by Histowiz.