Overview

  • Product name
    Anti-VEGF Receptor 2 antibody [EIC]
    See all VEGF Receptor 2 primary antibodies
  • Description
    Mouse monoclonal [EIC] to VEGF Receptor 2
  • Host species
    Mouse
  • Specificity
    The monoclonal antibody will detect native human VEGFR-2 / KDR in ELISA experiments and on the surface of different human cell types. It recognizes the transmembrane KDR e.g. in HUVECs which normally is fully glycosylated (200kDa form).
  • Tested applications
    Suitable for: ICC/IF, IHC-P, ELISA, IHC-Fr, IHC - Wholemount, Flow Cytmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Rat, Sheep, Human
  • Immunogen

    Recombinant fragment corresponding to Human VEGF Receptor 2 aa 20-757. Recombinant human soluble VEGF Receptor 2.
    Database link: P35968

  • Positive control
    • HUVEC cells. IHC-P: FFPE human placenta.
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab2349 as a replacement.

Applications

Our Abpromise guarantee covers the use of ab9530 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
ICC/IF Use at an assay dependent concentration. PubMed: 16763549
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ELISA Use a concentration of 1 - 5 µg/ml.
IHC-Fr Use a concentration of 1 µg/ml.
IHC - Wholemount Use at an assay dependent concentration.
Flow Cyt Use a concentration of 2 - 10 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

  • Application notes
    Is unsuitable for WB.
  • Target

    • Function
      Receptor for VEGF or VEGFC. Has a tyrosine-protein kinase activity. The VEGF-kinase ligand/receptor signaling system plays a key role in vascular development and regulation of vascular permeability. In case of HIV-1 infection, the interaction with extracellular viral Tat protein seems to enhance angiogenesis in Kaposi's sarcoma lesions.
    • Involvement in disease
      Defects in KDR are associated with susceptibility to hemangioma capillary infantile (HCI) [MIM:602089]. HCI are benign, highly proliferative lesions involving aberrant localized growth of capillary endothelium. They are the most common tumor of infancy, occurring in up to 10% of all births. Hemangiomas tend to appear shortly after birth and show rapid neonatal growth for up to 12 months characterized by endothelial hypercellularity and increased numbers of mast cells. This phase is followed by slow involution at a rate of about 10% per year and replacement by fibrofatty stroma.
    • Sequence similarities
      Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.
      Contains 7 Ig-like C2-type (immunoglobulin-like) domains.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications
      Phosphorylated. Dephosphorylated by PTPRB. Dephosphorylated by PTPRJ at Tyr-951, Tyr-996, Tyr-1054, Tyr-1059, Tyr-1175 and Tyr-1214.
    • Cellular localization
      Membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • CD309 antibody
      • CD309 antigen antibody
      • EC 2.7.10.1 antibody
      • Fetal liver kinase 1 antibody
      • FLK-1 antibody
      • FLK1 antibody
      • FLK1, mouse, homolog of antibody
      • Kdr antibody
      • Kinase insert domain receptor (a type III receptor tyrosine kinase) antibody
      • Kinase insert domain receptor antibody
      • KRD1 antibody
      • Ly73 antibody
      • Protein tyrosine kinase receptor FLK1 antibody
      • Protein-tyrosine kinase receptor flk-1 antibody
      • soluble VEGFR2 antibody
      • Tyrosine kinase growth factor receptor antibody
      • Vascular endothelial growth factor receptor 2 antibody
      • VEGFR 2 antibody
      • VEGFR antibody
      • VEGFR-2 antibody
      • VEGFR2 antibody
      • VGFR2_HUMAN antibody
      see all

    Images

    • Detection of VEGFR-2 extracellular domain of human umbilical vein endothelial cells (HUVEC) using monoclonal antibody VEGFR-2  (bold) versus control conditions.

    • IHC image of VEGF Receptor 2 staining in human placenta formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9530, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Immunocytochemical analysis of human aortic endothelial cells, labelling VEGF Receptor 2 with ab9530 (green). Cells cultured until 70% confluent and fixed with 4% paraformaldehyde for 15 minutes at room temperature. They were then incubated with ab9530, diluted in PBS containing 5% normal goat serum and 0.01% Triton X-100 at 4°C overnight. DAPI used to stain nuclei. 

    • ab9530 staining Human aortic endothelial cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 5% serum for 20 minutes at 22°C. Samples were incubated with primary antibody (1/2000 in PBS + 0.1% Triton-X 100 + 5% Goat serum) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody. Green - VEGF Receptor 2, Red - alpha tubulin, Blue - nuclear marker (DAPI).

      See Abreview

    • Wholemount Immunohistochemical analysis of lung tissue from a 6 day postnatal mouse, labelling VEGF Receptor 2 with ab9530. ab9530 was diluted 1/500 in 3% BSA and 0.1% Triton 100 in PBS buffer, and incubated at 4°C for 12 hours. The secondary used was a polyclonal Alexa Fluor 488 Goat Anti-Mouse IgG at 1/1000.

      See Abreview

    • ab9530 staining rat lung cells by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with paraformaldehyde and  blocked in 5% normal goat serum for 1 hour at room temperature. Samples were incubated with primary antibody (dilution 1/50) for 14 hours at 4°C; the diution buffer was 5% NGS in 1 X TBS-T (0.1%tween20). A Cy3® -conjugated Goat polyclonal to mouse IgG, dilution 1/100, was used as secondary antibody.

      See Abreview

    References

    This product has been referenced in:
    • Zhang J  et al. Pericardial application as a new route for implanting stem-cell cardiospheres to treat myocardial infarction. J Physiol 596:2037-2054 (2018). Read more (PubMed: 29736937) »
    • Zhao W  et al. ILK promotes angiogenic activity of mesenchymal stem cells in multiple myeloma. Oncol Lett 16:1101-1106 (2018). Read more (PubMed: 29963187) »
    See all 22 Publications for this product

    Customer reviews and Q&As

    1-10 of 36 Abreviews or Q&A

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 37°C
    Sample
    Cow Cell (Bovine Aortic Endothelial Cell)
    Specification
    Bovine Aortic Endothelial Cell
    Permeabilization
    Yes - 0.1% Triton X-100
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Feb 09 2015

    Application
    Flow Cytometry
    Fixation
    Paraformaldehyde
    Sample
    Human Cell (HUVEC, iPSC)
    Specification
    HUVEC, iPSC
    Permeabilization
    No

    Abcam user community

    Verified customer

    Submitted Jul 15 2014

    Application
    IHC - Wholemount
    Sample
    Mouse Tissue (Postnatal day 6 mouse lung)
    Specification
    Postnatal day 6 mouse lung

    Abcam user community

    Verified customer

    Submitted Dec 18 2013

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
    Antigen retrieval step
    None
    Sample
    Human Tissue sections (Human epithelial ovarian carcinoma tissue)
    Specification
    Human epithelial ovarian carcinoma tissue
    Permeabilization
    Yes - 0.2% Triton X-100
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 18 2013

    Application
    Immunocytochemistry
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 23°C
    Sample
    Mouse Cultured Cells (Mouse endothelial cells)
    Specification
    Mouse endothelial cells
    Permeabilization
    No
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 18 2013

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 22°C
    Sample
    Human Cell (Human aortic endothelial cells)
    Specification
    Human aortic endothelial cells
    Permeabilization
    Yes - 0.1% Triton X-100
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 17 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Sheep Cell (Pituitary)
    Specification
    Pituitary
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Feb 25 2013

    Answer

    Thank you for your reply. I have now returned to the office an hope I can help further with your enquiry.

    I can now confirm that of the antibodies we have directed against VEGF Receptor 2 (that are not raised in rabbit), most likely to react with the sheep protein are the mouse monoclonals ab10972, ab10973 and ab82266. This is based on the immunogen used to raise these antibodies. They have all be raised against the extracellular domain of the VEGF Receptor 2, however, this region covers the residues 1-745 and as I only have the region between 230-400 of the sheep protein to compare I cannot be at all certain that they will work, as you have observed with ab10972. We have ourselves not tested any of these antibodies with samples from sheep and therefore cannot guarantee their performance.

    If you would like, I can offer a testing discount with ab10973 or ab82266. More information on this offer can be found from the following link:

    www.abcam.com/abtrial

    However, I would suggest the rabbit antibodies I suggested, ab39521, ab62697, ab63613 and ab131510 (especially ab131510 as it has been used in ICC/IF as well as staining of paraffin embedded sections) would be more likely to work with your samples.

    Again, if you would like to participate in the Abtrial with any of these antibodies please do let me know.

    I hope this information has been of some help. If I can be of any further assistance, please do not hesitate to let me know.

    Read More

    Answer

    Thank you for your reply. I have now checked through the antibodies we have directed against VEGF Receptor 2 and the likelihood that any of them will be able to cross react with the protein in your sheep samples.

    The two fragments of sequence from SwissProt (http://www.uniprot.org/uniprot/Q8MI25 and http://www.uniprot.org/uniprot/Q8MI23) align to ˜230-400 and 1020-1200 of human and mouse VEGF Receptor 2 respectively. From this, I have been able to identify a total of 4 antibodies that would be likely to cross react. These are ab39521, ab62697, ab63613 and ab131510.

    Of these I think the antibody ab131510 would be most worthwhile to try as this has been used in ICC/IF as well as staining of paraffin embedded sections.

    As discussed over the phone, we would not be able to guarantee the antibody will cross react. This was also the case with ab9530 that you tried. Unless the species and application you wish to use are listed on the datasheet of the product then they would not be covered by the Abpromise guaranteed. I am sorry that this was not made clear to you. The terms of the Abprimise can be found from the following link:

    www.abcam.com/abpromise

    If a species or application you wish to use has not been tested with the antibody you want to use I would encourage you to contact us as we may be able to advise if it is worthwhile trying it. You may also be eligible for the Abtrial discount scheme. More details of this scheme can be found from the following link:

    www.abcam.com/abtrial

    As discussed, I would be willing to provide you with ab131510 for you free of charge if you would be interested as a gesture of goodwill. In return, I would appreciate it if you would provide an Abreview once you have used the antibody. This should only take 5-10 minutes to complete.

    I cannot provide you with ab10972 as initially discussed as I would not be as sure of its reactivity due to the immunogen used to raise the antibody not being within the fragments we have for the sheep protein.

    I hope this information has been of help. I look forward to hearing how you would like to proceed.

    Read More

    Answer

    Thank you for your enquiry and interest.

    In order to be able to find the suitable isotype control, it is important to know the isotype of these antibodies are:

    ab9530 - Mouse IgG1

    ab27619 - Rabbit IgG (not determined further)

    You can use our advanced search engine to find the most appropriate controls:

    https://www.abcam.com/index.html?c=718

    If you need any further assistance in the future, please do not hesitate to contact me.

    Read More

    1-10 of 36 Abreviews or Q&A

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