Recombinant
RabMAb

Anti-YAP1 (phospho S127) antibody [EP1675Y] (ab76252)

Overview

  • Product name
    Anti-YAP1 (phospho S127) antibody [EP1675Y]
    See all YAP1 primary antibodies
  • Description
    Rabbit monoclonal [EP1675Y] to YAP1 (phospho S127)
  • Specificity
    ab76252 detects YAP1 phosphorylated at Serine 127.
  • Tested applications
    Suitable for: WB, IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human YAP1 (phospho S127).

  • Positive control
    • WB: HeLa, C6 and NIH/3T3 cell lysates treated with FBS + Calyculin A. IHC-P: Mouse kidney, rat stomach and human adenocarcinoma of the endometrium tissues.
  • General notes

    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 ab76252 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 1/10000 - 1/50000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 - 75 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

  • Application notes
    Is unsuitable for ICC/IF.
  • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of endometrium tissue labelling YAP1 (phospho S127) with purified ab76252 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • All lanes : Anti-YAP1 (phospho S127) antibody [EP1675Y] (ab76252) at 1/1000 dilution

      Lane 1 : 293A grown in serum-free media overnight, whole cell lysate
      Lane 2 : 293A grown in serum-free media overnight, then 10% FBS was added to medium for 1 hour, whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size : 65 - 75 kDa
      Observed band size : 75 kDa (why is the actual band size different from the predicted?)


      Exposure time : 3 minutes

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling YAP1 (phospho S127) with purified ab76252 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • All lanes : Anti-YAP1 (phospho S127) antibody [EP1675Y] (ab76252) at 1/50000 dilution (purified)

      Lane 1 : HeLa cell lysate - untreated
      Lane 2 : HeLa cell lysate - treated with Calyculin A

      Lysates/proteins at 10 µg per lane.

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 65 - 75 kDa
      Observed band size : 65 - 75 kDa

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue labelling YAP1 (phospho S127) with purified ab76252 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • All lanes : Anti-YAP1 (phospho S127) antibody [EP1675Y] (ab76252) at 1/5000 dilution (unpurified)

      Lane 1 : C6 treated with Calyculin A
      Lane 2 : NIH/3T3 treated with Calyculin A

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size : 65 - 75 kDa
      Observed band size : 65 - 75 kDa
    • Dot blot analysis of YAP1 (pS127) peptide (Lane 1) and YAP1 non-phospho peptide (Lane 2) labelling YAP1 (pS127) with ab76252 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    References

    This product has been referenced in:
    • Wu J  et al. Inhibitory effect and mechanism of exogenous mammalian sterile 20-like kinase 1 on the growth of human colorectal cancer. Oncol Lett 13:2656-2664 (2017). WB ; Human . Read more (PubMed: 28454447) »
    • Zhang X  et al. The essential role of YAP O-GlcNAcylation in high-glucose-stimulated liver tumorigenesis. Nat Commun 8:15280 (2017). WB, IF ; Human . Read more (PubMed: 28474680) »

    See all 7 Publications for this product

    Customer reviews and Q&As

    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% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Dec 21 2016

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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