Anti-Glucocorticoid Receptor antibody (ab3578)

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5 μm sections were cut and mounted on saline-coated slides, dried, deparaffinized in xylene, and rehydrated in alcohol. Endogenous peroxidase was quenched by 3% hydrogen peroxidase for 10 minutes followed by heat antigen retrieval in 0.01 M citrate buffer, pH 6.0, in a microwave oven at a medium oven setting. Slides were incubated overnight at 4°C with mouse monoclonal [3D5] to glucocorticoid receptor (ab9568) (Abcam, UK) in 1 : 100 dilution. Immunoreaction was detected and visualized by using a DakoCytomation LSAB2 System (HRP code K0673). Positive cases were determined by exhibiting a focal or diffuse moderate to dark brown nuclear staining pattern in more than 10% of the cells. The percentage of cases showing positive GR expression was calculated for various categories of mammary epithelium.

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