Androgen Receptor Antibodies

Immunofluorescent analysis of androgen receptor (green) in LNCaP (right panel) and negative control U2OS cells (left panel). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Cat. No. 37525) for 15 minutes at room temperature. Cells were probed with an androgen receptor monoclonal antibody (Cat. No. MA5-13426) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Cat. No. 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight-554 Phalloidin (Cat. No. 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Cat. No. 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Immunohistochemistry analysis of androgen receptor showing staining in the cytoplasm and nucleus of paraffin-treated human prostate tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with an androgen receptor polyclonal antibody (Cat. No. PA1-110) diluted by 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.