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Cardiovascular Angiogenesis Growth Factors VEGF VEGF Receptors
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Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)

  • Datasheet
  • SDS
Reviews (10)Q&A (2)References (26)

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Immunocytochemistry/ Immunofluorescence - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)
  • Western blot - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)

Key features and details

  • Rabbit polyclonal to VEGF Receptor 2 (phospho Y1054 + Y1059)
  • Suitable for: ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

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Protein
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Primary
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Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

Overview

  • Product name

    Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody
    See all VEGF Receptor 2 primary antibodies
  • Description

    Rabbit polyclonal to VEGF Receptor 2 (phospho Y1054 + Y1059)
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic phospho peptide (Human)containing Tyrosines 1054 and 1059. This sequence is conserved in mouse and rat.

  • Positive control

    • IF: VEGF treated HUVEC whole cells. WB: VEGF treated HUVEC, MDA-MB-231 and MCF7 whole cell extract.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.30
    Preservative: 0.05% Sodium azide
    Constituents: PBS, 0.1% BSA, 50% Glycerol
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated VEGFR 2. The final product is generated by affinity chromatography using a VEGFR 2 derived peptide that is phosphorylated at Tyrosines 1054 and 1059.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Growth Factors
    • VEGF
    • VEGF Receptors
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Signal Transduction
    • Growth Factors/Hormones
    • VEGF
    • Stem Cells
    • Hematopoietic Progenitors
    • Surface Molecules
    • Cancer
    • Growth factors
    • VEGF
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Growth factor receptors
    • Stem Cells
    • Endothelial Progenitors
    • Endothelial Markers
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Cardiovascular
    • Cardiovascular Markers
    • Cell Markers
    • Endothelial Cells
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human VEGF Receptor 2 protein (Active) (ab106310)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab5473 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
WB
Human
All applications
Mouse
Rat
Application Abreviews Notes
ICC/IF (2)
1/250.
WB (1)
Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 200 kDa.
Notes
ICC/IF
1/250.
WB
Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 200 kDa.

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.
  • Target information above from: UniProt accession P35968 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3791 Human
    • Entrez Gene: 16542 Mouse
    • Entrez Gene: 25589 Rat
    • Omim: 191306 Human
    • SwissProt: P35968 Human
    • SwissProt: P35918 Mouse
    • SwissProt: O08775 Rat
    • Unigene: 479756 Human
    • Unigene: 285 Mouse
    • Unigene: 88869 Rat
    see all
  • 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

  • Immunocytochemistry/ Immunofluorescence - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)
    Immunocytochemistry/ Immunofluorescence - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)

    Immunofluorescence analysis of HUVEC cells labelling VEGF Receptor 2 (phospho Y1054 + Y1059) (PAnel a: green) using ab5473 at 2 ug/ml in 0.1% BSA for 3 hours at room temperature, follwed by Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal at 1/2000 dilution. Pbel b: Nuclei stained with DAPI (blue), Panel c: F-actin stained with Alexa Fluor® 555 Rhodamine Phalloidin (red), Panel d: Merged images. The images were captured at 60X magnification.

    Prior antibody incubation, HUVEC (Human umbilical vein endothelial cell line) cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes and blocked with 1% BSA for 1 hour at room temperature, followed by treatment with 100 ng of VEGF for 30 minutes. Assay was done on 90% confluent log phase HUVEC cells.

     

  • Western blot - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)
    Western blot - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)
    All lanes : Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473) at 1/1000 dilution

    Lane 1 : HUVEC (Human umbilical vein endothelial cell line) whole cell extract with Skimmed milk
    Lane 2 : HUVEC cells treated with 25ng/ml VEGF for 5 minutes with Skimmed milk
    Lane 3 : MDA-MB-231 (human breast adenocarcinoma cell line) whole cell extract with Skimmed milk
    Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell extract with Skimmed milk

    Lysates/proteins at 30 µg per lane.

    Blocking peptides at 5 % per lane.

    Secondary
    All lanes : Goat anti-rabbit IgG HRP conjugate at 1/5000 dilution


    A 130 kDa band corresponding to VEGFR (pYpY1054/1059) was observed across cell lines tested.

    Protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard.

    Resolved proteins were then transferred onto a nitrocellulose membrane by over night wet transfer. 

    Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.

     

Protocols

  • Peptide Competition Experiment
  • Western blotting protocol

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (26)

    Publishing research using ab5473? Please let us know so that we can cite the reference in this datasheet.

    ab5473 has been referenced in 26 publications.

    • Li L  et al. Brivanib, a multitargeted small-molecule tyrosine kinase inhibitor, suppresses laser-induced CNV in a mouse model of neovascular AMD. J Cell Physiol 235:1259-1273 (2020). PubMed: 31270802
    • Cho YR  et al. A novel role for a-viniferin in suppressing angiogenesis by blocking the VEGFR-2/p70S6K signaling pathway. Phytother Res N/A:N/A (2020). PubMed: 32400050
    • Jin B  et al. Yangjing Capsule Can Improve the Function of the Testicular Angiogenesis through Activating VEGFA/eNOS Signaling Pathway. Evid Based Complement Alternat Med 2020:1957267 (2020). PubMed: 32382279
    • Zhang Q  et al. Acidic Bile Salts Induce Epithelial to Mesenchymal Transition via VEGF Signaling in Non-Neoplastic Barrett's Cells. Gastroenterology 156:130-144.e10 (2019). PubMed: 30268789
    • Zhang Q  et al. Improved Antitumor Efficacy of Combined Vaccine Based on the Induced HUVECs and DC-CT26 Against Colorectal Carcinoma. Cells 8:N/A (2019). PubMed: 31121964
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Filter by Application

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    1-10 of 10 Abreviews

    Western blot abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Cardiac)
    Gel Running Conditions
    Reduced Denaturing (8%)
    Loading amount
    40 µg
    Specification
    Cardiac
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 03 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Brain cell culture)
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Brain cell culture
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 21 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA Buffer, pH 9.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Liver
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 11 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Pig Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA Buffer, pH 9.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Liver
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 11 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA Buffer, pH 9.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Liver
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 11 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Kidney H9 Organoids)
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney H9 Organoids
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 11 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA Buffer, pH 9.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Liver
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 11 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA Buffer, pH 9.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Liver
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 11 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Heart)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH6.0
    Permeabilization
    No
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Mengcheng Shen

    Verified customer

    Submitted Dec 20 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate bufer
    Sample
    Rat Tissue sections (rat bone marrow tissue)
    Specification
    rat bone marrow tissue
    Permeabilization
    No
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 07 2014

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