Alexa Fluor® 647 Anti-Progesterone Receptor antibody [SP42]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Progesterone Receptor antibody [SP42] (AB270198)

Flow cytometry overlay histogram showing T47D cells stained with ab270198 (red line). The cells were fixed with 4 % formaldehyde (10 min) and then permeabilized with 0.1 % PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab270198) (1x 10⁶ in 100μl at 0.04 μg/ml (1/12500)) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor^® 647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 40 mW Red laser (640nm) and 660/20 bandpass filter.

This antibody gave a positive signal in T47D cells fixed with 80 % methanol (5 min) / permeabilized with 0.1 % PBS-Triton X-100 for 15 min used under the same conditions.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Progesterone Receptor antibody [SP42] (AB270198)

Immunofluorescence staining of Progesterone Receptor in T47D cells using ab270198. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab270198 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in T47D cells fixed with 100% methanol (5 min).