Anti-YAP1 antibody [EP1674Y]

• IP

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Immunoprecipitation - Anti-YAP1 antibody [EP1674Y] (AB52771)

ab52771 (purified) at 1 : 20 dilution (0.5 μg) immunoprecipitating YAP1 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.
Lane 1 : HeLa whole cell lysate 10 μg input.
Lane 2 : ab52771 in HeLa whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab52771 in HeLa whole cell lysate.

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.

Blocking/Diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-YAP1 antibody [EP1674Y] (ab52771)

Predicted band size: 54 kDa

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• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-YAP1 antibody [EP1674Y] (AB52771)

Immunocytochemistry/Immunofluorescence analysis of MCF-7 (Human breast adenocarcinoma cell line) cells labeling YAP1 with purified ab52771 at 1/500.

Cells were fixed with 4% paraformaldehyde and permeabilized by 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077).

Nuclei counterstained with DAPI (blue).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-YAP1 antibody [EP1674Y] (AB52771)

Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with unpurified ab52771 (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 (ab52771, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) (1 μg/1x10⁶ cells) used under the same conditions.

Acquisition of >5,000 events was performed.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-YAP1 antibody [EP1674Y] (AB52771)

Immunofluorescence revealed that YAP1 expression was significantly lower in iVSD hearts compared to control hearts.

YAP1 (green), and DAPI (blue) staining are shown; Scale bar = 25 μm.

Cryosections of human heart tissue were blocked using 10% FBS for 30 min, and then incubated with anti YAP1 (ab52771, 1 : 200) at room temperature for 2 h. The slides were then incubated with the following secondary antibody, Alexa Fluor^® 488-conjugated anti-rabbit (Abcam, ab150073; 1 : 1,000 dilution). Cell nuclei were stained with DAPI.

Ye et al PLoS One. 2015 Oct 1;10(10):e0139712. doi: 10.1371/journal.pone.0139712. eCollection 2015. Fig 2. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YAP1 antibody [EP1674Y] (AB52771)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling YAP1 with purified ab52771 at 1 : 50 dilution (1.78 μg/ml).

Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody.

PBS instead of the primary antibody was used as the negative control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-YAP1 antibody [EP1674Y] (AB52771)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling YAP1 with purified ab52771 at 1/20 dilution (10μg/ml) (red).

Cells were fixed with 4% paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

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Western blot - Anti-YAP1 antibody [EP1674Y] (AB52771)

Blocking/Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-YAP1 antibody [EP1674Y] (ab52771) at 1/5000 dilution

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates 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: 54 kDa

Observed band size: 72 kDa

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• WB

AbReview20683****

Western blot - Anti-YAP1 antibody [EP1674Y] (AB52771)

All lanes are unpurified ab52771 at a 1/1000 dilution plus whole cell lysate prepared from U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells, treated with DMSO or 1 μM and 10 μM LY294002, 50 μg positive control loaded.

Primary antibody was incubated for 16 hours at 4°C.

Blocking step was performed using 5% milk for 1 hour at 23°C.

Secondary used was a goat polyclonal conjugated to HRP, at a 1/10,000 dilution.

All lanes:

Western blot - Anti-YAP1 antibody [EP1674Y] (ab52771)

Predicted band size: 54 kDa

Observed band size: 70 kDa

true

Exposure time: 1min

Image courtesy of an anonymous Abreview.

• WB

Unknown

Western blot - Anti-YAP1 antibody [EP1674Y] (AB52771)

All lanes:

Western blot - Anti-YAP1 antibody [EP1674Y] (ab52771) at 1/50000 dilution

All lanes:

HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 54 kDa

Observed band size: 72 kDa

false

• WB

Unknown

Western blot - Anti-YAP1 antibody [EP1674Y] (AB52771)

Blocking/Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-YAP1 antibody [EP1674Y] (ab52771) at 1/50000 dilution

All lanes:

Human thyroid cancer lysates at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 54 kDa

Observed band size: 72 kDa

false