Recombinant Anti-Progesterone Receptor antibody [SP2] (ab16661)

Immunocytochemistry/ Immunofluorescence analysis of T-47D (human ductal breast epithelial tumor epithelial cell) cells labeling Progesterone Receptor with purified ab16661 at 1/100 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Flow Cytometry analysis of T-47D (human ductal breast epithelial tumor epithelial cell) cells labeling Progesterone Receptor with purified ab16661 at 1/220 dilution (1.04 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue.

Immunohistochemistry analysis of human breast carcinoma tissue labelling SP2 with ab16661.

ab16661 staining rat ovary tissue sections by IHC-P.  Sections were fixed in 10% buffered formalin and subjected to heat mediated antigen retrieval in 10mM citrate buffer pH 6.0 prior to blocking with 5% serum for 20 minutes at 20°C.  The primary antibody was diluted 1/50 in TBS (with Tween) and incubated with the sample for 16 hours at 4°C.  A HRP conjugated goat anti-rabbit was used as the secondary antibody.

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Overlay histogram showing T47D cells stained with ab16661 (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 (ab16661, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.