Anti-Androgen Receptor antibody [AR 441] (ab9474)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Androgen Receptor antibody [AR 441] (ab9474)

ab9474 staining human prostate by IHC-P.

Immunocytochemistry/ Immunofluorescence - Androgen Receptor antibody [AR 441] (ab9474)

ab9474 staining the Androgen Receptor in the human 22Rv1 Prostate Cancer Cell Line by ICC/IF (immunocytochemistry/immunofluorescence). Cells were formaldehyde fixed, permeabilized by 0.1% Triton for 15 minutes, and blocked with glycine (7.5mg/ml) for 15 minutes at room temperature. The sample was incubated with the primary antibody (1/50 in PBS) for 12 hours at 4ºC. An undiluted Alexa Fluor 488^®-conjugated Donkey anti-mouse polyclonal was used as the secondary.

This image is courtesy of an Abreview submitted by Dr Victor Campa

Flow Cytometry - Androgen Receptor antibody [AR 441] (ab9474)

Overlay histogram showing MCF-7 cells stained with ab9474 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9474, 1/10 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight ® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Flow Cytometry-Androgen Receptor antibody [AR 441](ab9474)

Overlay histogram showing PC3 cells stained with ab9474 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9474, 1/10 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight ® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in PC3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.