Overview

  • Product name

    Anti-VAV1 (phospho Y174) antibody [EP510Y]
    See all VAV1 primary antibodies
  • Description

    Rabbit monoclonal [EP510Y] to VAV1 (phospho Y174)
  • Host species

    Rabbit
  • Specificity

    This antibody detects VAV1 phosphorylated at Tyrosine 174.
  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, Dot blotmore details
    Unsuitable for: IHC-P or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human VAV1 (phospho Y174). The exact sequence is proprietary.

  • Positive control

    • WB: Jurkat cell lysate treated with pervanadate.
  • General notes

    Mouse, 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76225 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 1/1000. Detects a band of approximately 101 kDa (predicted molecular weight: 101 kDa).
Flow Cyt Use at an assay dependent concentration.
ICC/IF 1/100 - 1/500.
Dot blot Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-P or IP.
  • Target

    • Function

      Couples tyrosine kinase signals with the activation of the Rho/Rac GTPases, thus leading to cell differentiation and/or proliferation.
    • Tissue specificity

      Widely expressed in hematopoietic cells but not in other cell types.
    • Sequence similarities

      Contains 1 CH (calponin-homology) domain.
      Contains 1 DH (DBL-homology) domain.
      Contains 1 PH domain.
      Contains 1 phorbol-ester/DAG-type zinc finger.
      Contains 1 SH2 domain.
      Contains 2 SH3 domains.
    • Domain

      The DH domain is involved in interaction with CCPG1.
    • Post-translational
      modifications

      Phosphorylated on tyrosine residues.
    • Information by UniProt
    • Database links

    • Alternative names

      • Oncogene vav antibody
      • p95Vav antibody
      • Proto-oncogene vav antibody
      • Protooncogene vav antibody
      • VAV 1 antibody
      • VAV 1 oncogene antibody
      • VAV antibody
      • Vav proto oncogene antibody
      • VAV_HUMAN antibody
      • VAV1 antibody
      see all

    Images

    • All lanes : Anti-VAV1 (phospho Y174) antibody [EP510Y] (ab76225) at 1/1000 dilution

      Lane 1 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
      Lane 2 : Jurkat treated with 50mM pervanadate for 5minutes whole cell lysate
      Lane 3 : Jurkat treated with 50mM pervanadate for 5min whole cell lysate 10 µg. Then the membrane was incubated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 101 kDa



      Blocking and diluting buffer: 5% NFDM/TBST

    • Immunocytochemistry/Immunofluorescence analysis of Untreated Jurkat cells and Pervanadate (1mM,30min) treated Jurkat cells labelling VAV1 (phospho Y174) with purified ab76225 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100, and counterstained with ab7291 anti-Tubulin (mouse mAb) 1:1000 (1ug/ml). An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

      Negative Control 1: Rabbit primary antibody and anti-mouse secondary antibody(ab150120)

      Negative Control 2: Mouse primary antibody(ab7291) and anti-rabbit secondary antibody(ab150077)

    • Flow Cytometry analysis of Jurkat (human acute T cell leukemia) untreated/treated with 1mM pervanadate for 30min cells labeling VAV1 with unpurified ab76225 at 1/200 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. Untreated control (Green).

    • Dot blot analysis of VAV1 (pY174) peptide (Lane 1) and VAV1 non-phospho peptide (Lane 2) labelling VAV1 (phospho Y174) with ab76225 at a dilution of 1/1000. A Peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    References

    This product has been referenced in:

    • Zhang C  et al. CD100-plexin-B1 induces epithelial-mesenchymal transition of head and neck squamous cell carcinoma and promotes metastasis. Cancer Lett 455:1-13 (2019). Read more (PubMed: 30981760) »
    • Abate F  et al. Activating mutations and translocations in the guanine exchange factor VAV1 in peripheral T-cell lymphomas. Proc Natl Acad Sci U S A 114:764-769 (2017). Read more (PubMed: 28062691) »
    See all 9 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Application
    Western blot
    Loading amount
    12 µg
    Gel Running Conditions
    Reduced Denaturing (gel 8%)
    Sample
    Human Cell lysate - whole cell (SKOV3 cells were transfected with VAV1)
    Specification
    SKOV3 cells were transfected with VAV1
    Blocking step
    Blocking one (nacalai tesque) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 27°C

    Abcam user community

    Verified customer

    Submitted Jul 18 2013

    Answer

    The difference between these two products is that ab76225 is a rabbit monoclonal antibody raised while ab47282 is a rabbit polyclonal both of these antibodies were raised against the corresponding residues around the phophorylated site at the 174 amino acid in the Vav protein sequence. Unfortunately neither of these antibodies have been tested in flow cytometry experiments, thus we do not know if they will work or not. Either one of these antibodies is likely to work in flow cytometery since they are recognizing the proper tertiary structure of the protein as evidenced by the fact that the work in immunocytochemistry experiments. If you would like we can provide you with a testing discount for one or both of these antibodies to test in flow cytometry. In order to receive this offer you should inform us that you would like to purchase this antibody for testing in flow cytometry and we will provide you with a testing discount code. Then, you would buy the antibody at full price and use it with your samples. After testing the product with your samples you would then report your results to us through an Abreview on our website within 120 days of your purchase. Upon submitting the review your testing discount code would be activated and would be worth the price of an antibody with the same or lesser value of the original product. The test code must be redeemed within 120 days of purchase of the original product.

    Read More

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