Anti-Progesterone Receptor antibody [YR85]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (AB32085)

Immunofluorescence staining of Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085) in T47D cells (+ve control) and HepG2 cells (-ve control). The cells were fixed with 4% paraformaldehyde (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 ab32085 at 0.2 μg/mL dilution and ab7291, Anti-alpha Tubulin antibody [DM1A], at 1/1000 dilution, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150087) at 1/1000 (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with the Perkin Elmer Operetta and a maximum intensity projection of confocal planes is shown.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (AB32085)

Immunofluorescence staining of Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085) in T47D cells (+ve control) and HepG2 cells (-ve control). The cells were fixed with 100% Methanol (5 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 ab32085 at 0.2 μg/mL dilution and ab7291, Anti-alpha Tubulin antibody [DM1A], at 1/1000 dilution, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150087) at 1/1000 (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with the Perkin Elmer Operetta and a maximum intensity projection of confocal planes is shown.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (AB32085)

ICC/IF image of ab32085 stained DU145 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab32085 at 5μg/ml overnight at +4°C. The secondary antibody (green) was DyLight^® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progesterone Receptor antibody [YR85] (AB32085)

Immunohistochemical analysis of progesterone receptor expression in paraffin embedded human breast carcinoma, using 1/100 ab32085.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Progesterone Receptor antibody [YR85] (AB32085)

Intracellular Flow Cytometry analysis of T47D (human mammary gland ductal carcinoma) cells labeling Progesterone Receptor with purified ab32085 at 1/1100 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• WB

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Western blot - Anti-Progesterone Receptor antibody [YR85] (AB32085)

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085) at 1/2000 dilution

All lanes:

T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 98 kDa

Observed band size: 118 kDa

false

Exposure time: 40s

• WB

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Western blot - Anti-Progesterone Receptor antibody [YR85] (AB32085)

All lanes:

Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085) at 1/1000 dilution

All lanes:

Human breast cancer lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Predicted band size: 98 kDa

Observed band size: 118 kDa

false

Exposure time: 180s

• OI-RD Scanning

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OI-RD Scanning - Anti-Progesterone Receptor antibody [YR85] (AB32085)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.