Anti-Progesterone Receptor antibody [Alpha PR6]

4 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progesterone Receptor antibody [Alpha PR6] (ab2765)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) was performed on human uterus tissue. Antigen retrieval was performed using 10mM sodium citrate followed by microwave treatment for 8-15 minutes. Endogenous peroxidases were blocked in 3% H202-methanol for 15 minutes and tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with ab2765 (1:20) overnight in a humidified chamber. Tissues were washed in PBST and detection was performed using a secondary antibody conjugated to HRP. DAB staining buffer was applied and tissues were counterstained with hematoxylin and prepped for mounting. Images were taken at 40X magnification.

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  Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [Alpha PR6] (ab2765)

  Immunocytochemistry/ Immunofluorescence analysis of T47D cells untreated (left) or stimulated with 100nm promegestone for 1 hour (right), labeling Progesterone Receptor with ab2765 (green). The cells were fixed with formalin for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 3% Blocker BSA for 15 minutes at room temperature. Cells were stained with Anti-Progesterone Receptor antibody [Alpha PR6] - ChIP Grade (ab2765) at a dilution of 1/100 for 1 hour at 37C, and then incubated with a Alexa Fluor 488 goat anti-mouse IgG secondary antibody at a dilution of 1/1000 for 30 minutes at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye.

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  Flow Cytometry - Anti-Progesterone Receptor antibody [Alpha PR6] (ab2765)

  Overlay histogram showing T47D cells stained with ab2765 (red line). The cells were fixed with 80% 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 (ab2765, 0.5µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (Mouse IgG2a, Kappa Monoclonal [B12/8] - Isotype Control ab91361, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  

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  Western blot - Anti-Progesterone Receptor antibody [Alpha PR6] (ab2765)

  All lanes: Western blot - Anti-Progesterone Receptor antibody [Alpha PR6] (ab2765) at 1 µg/mL

  Lane 1: T47D cell lysate untreated (-) at 20 µg

  Lane 2: T47D cell lysate stimulated (+) with 100 nm promegestone for 1 hour at 20 µg

  Secondary

  All lanes: Goat anti-Mouse IgG-HRP at 1/2000 dilution

  Predicted band size: 98 kDa