Recombinant
RabMAb

Anti-YAP1 antibody [EP1674Y] (ab52771)

Overview

  • Product name
    Anti-YAP1 antibody [EP1674Y]
    See all YAP1 primary antibodies
  • Description
    Rabbit monoclonal [EP1674Y] to YAP1
  • Host species
    Rabbit
  • Specificity
    This antibody reacts with YAP1.
  • Tested applications
    Suitable for: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human YAP1 aa 350-450 (C terminal).
    (Peptide available as ab173007)

  • Positive control
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52771 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 65 kDa).
IP 1/20.

For unpurified use at 1:70. 

Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1:50.

 

IHC-P 1/50.
ICC/IF 1/100 - 1/500.

Target

  • Function
    Transcriptional regulator which can act both as a coactivator and a corepressor and is the critical downstream regulatory target in the Hippo signaling pathway that plays a pivotal role in organ size control and tumor suppression by restricting proliferation and promoting apoptosis. The core of this pathway is composed of a kinase cascade wherein MST1/MST2, in complex with its regulatory protein SAV1, phosphorylates and activates LATS1/2 in complex with its regulatory protein MOB1, which in turn phosphorylates and inactivates YAP1 oncoprotein and WWTR1/TAZ. Plays a key role to control cell proliferation in response to cell contact. Phosphorylation of YAP1 by LATS1/2 inhibits its translocation into the nucleus to regulate cellular genes important for cell proliferation, cell death, and cell migration. The presence of TEAD transcription factors are required for it to stimulate gene expression, cell growth, anchorage-independent growth, and epithelial mesenchymal transition (EMT) induction. Isoform 2 and isoform 3 can activate the C-terminal fragment (CTF) of ERBB4 (isoform 3).
  • Tissue specificity
    Increased expression seen in some liver and prostate cancers. Isoforms lacking the transactivation domain found in striatal neurons of patients with Huntington disease (at protein level).
  • Sequence similarities
    Belongs to the YORKIE family.
    Contains 2 WW domains.
  • Post-translational
    modifications
    Phosphorylated by LATS1 and LATS2; leading to cytoplasmic translocation and inactivation. Phosphorylated by ABL1; leading to YAP1 stabilization, enhanced interaction with TP73 and recruitment onto proapoptotic genes; in response to DNA damage.
  • Cellular localization
    Cytoplasm. Nucleus. Both phosphorylation and cell density can regulate its subcellular localization. Phosphorylation sequesters it in the cytoplasm by inhibiting its translocation into the nucleus. At low density, predominantly nuclear and is translocated to the cytoplasm at high density.
  • Information by UniProt
  • Database links
  • Alternative names
    • 65 kDa Yes associated protein antibody
    • 65 kDa Yes-associated protein antibody
    • COB1 antibody
    • YAp 1 antibody
    • YAP 65 antibody
    • YAP antibody
    • YAP1 antibody
    • YAP1_HUMAN antibody
    • YAP2 antibody
    • YAP65 antibody
    • yes -associated protein delta antibody
    • Yes associated protein 1 65kDa antibody
    • Yes associated protein 1 antibody
    • Yes associated protein 2 antibody
    • yes associated protein beta antibody
    • YKI antibody
    • Yorkie homolog antibody
    see all

Images

  • ab52771 (purified) at 1:20 dilution (0.5µg) immunoprecipitating YAP1 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
    Lane 2 (+): ab52771 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52771 in HeLa whole cell lysate
    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling YAP1 with purified ab52771 at 1:20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 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).
  • 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 Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
  • Anti-YAP1 antibody [EP1674Y] (ab52771) at 1/50000 dilution (purified) + Human thyroid cancer lysates at 15 µg

    Secondary
    Rabbit monoclonal [EP1674Y] to YAP1 (ab52771) at 1/2000 dilution (Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG)

    Predicted band size: 65 kDa



    Blocking and diluting buffer : 5% NFDM/TBST
  • Anti-YAP1 antibody [EP1674Y] (ab52771) at 1/5000 dilution (purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 65 kDa



    Blocking and diluting buffer : 5% NFDM/TBST
  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling YAP1 with purified ab52771 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised by 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

  • Immunohistochemical staining of YAP1 in paraffin embedded human breast carcinoma tissue using unpurified ab52771 at a 1/100 dilution.

  • Anti-YAP1 antibody [EP1674Y] (ab52771) at 1/50000 dilution (unpurified) + Hela cell lysate at 10 µg

    Secondary
    Goat anti-rabbit HRP labeled at 1/2000 dilution

    Predicted band size: 65 kDa
    Observed band size: 65 kDa

  • Overlay histogram showing HeLa 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) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes are unpurified ab52771 at a 1/1000 dilution plus whole cell lysate prepared from U87MG tumour cell line, treated with DMSO or 1uM &10uM LY294002, 50µg 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.

    See Abreview

References

This product has been referenced in:
  • Kang W  et al. miR-375 is involved in Hippo pathway by targeting YAP1/TEAD4-CTGF axis in gastric carcinogenesis. Cell Death Dis 9:92 (2018). WB, IP . Read more (PubMed: 29367737) »
  • Cho YS  et al. Regulation of Yki/Yap subcellular localization and Hpo signaling by a nuclear kinase PRP4K. Nat Commun 9:1657 (2018). Read more (PubMed: 29695716) »

See all 41 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (tumor xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
tumor xenograft
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
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Submitted Jan 02 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin carcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
skin carcinoma
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% buffered normal formalin
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Submitted Nov 27 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
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Submitted Nov 27 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (293A)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
10 µg
Specification
293A
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Submitted Dec 21 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Xenopus laevis Cell lysate - whole cell (oocyte)
Total protein in input
1500 µg
Immuno-precipitation step
Other - Dynabeads Protein A
Specification
oocyte
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Submitted Oct 27 2016

Application
Western blot
Sample
Xenopus laevis Cell lysate - whole cell (oocyte)
Gel Running Conditions
Reduced Denaturing (7,5)
Loading amount
20 µg
Specification
oocyte
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Oct 14 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (H22 (Hepatoma cell line))
Permeabilization
No
Specification
H22 (Hepatoma cell line)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Submitted Mar 18 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (BEL-7402)
Permeabilization
No
Specification
BEL-7402
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Submitted Mar 15 2016

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (BEL-7402)
Total protein in input
80 µg
Treatment
80 µM Doxorubicin for 24hrs
Immuno-precipitation step
Protein A/G
Specification
BEL-7402
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Submitted Feb 11 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (BEL-7402)
Gel Running Conditions
Non-reduced Denaturing (10%)
Loading amount
50 µg
Treatment
80 µM actinomycin for 24hrs
Specification
BEL-7402
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jan 29 2016

1-10 of 16 Abreviews or Q&A

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