Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

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Figure 1.  The Human Phospho-VEGF R3 DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western analysis
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Product Details
Citations (5)
Supplemental Products
Reviews (2)

Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
Cell lysates (100 µL)
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated human VEGF R3 / Flt-4 in cell lysates. An immobilized capture antibody specific for VEGF R3 / Flt-4 binds both phosphorylated and unphosphorylated VEGF R3 / Flt-4. After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

Product Features

  • Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Available in 2, 5, and 15-(96-well) plate pack sizes
  • Economical alternative to Western blot

Kit Content

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control

Other Reagents Required

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Scientific Data

Figure 1. The Human Phospho-VEGF R3 DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western analysis Lysates prepared from human VEGF R3 transfected NS0 cells (NS0-hVEGF R3) were diluted in a series and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an anti-VEGF R3 monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with a biotinylated anti-phosphotyrosine monoclonal antibody (Catalog # BAM1676) to detect phospho-VEGF R3. Bands were visualized with Streptavidin-HRP (Cat # DY998) followed by chemiluminescent detection using WesternGloTM Chemiluminescent Detection Substrate (Catalog # AR004). Human Phospho-VEGF R3 can be detected in this DuoSet IC ELISA by using approximately 10 to 15 times less lysate than is needed for a conventional IP-Western blot.

Figure 2. The specificity of the Human Phospho-VEGF R3 DuoSet IC ELISA is confirmed by receptor competition The indicated amounts of recombinant extracellular domains of human VEGF R3 (Catalog #349-F4), human VEGF R1 (Catalog #321-FL), human VEGF R2 (Catalog #357-KD) or human Tie-2 (Catalog #313-TI) were added to 20 μg NS0-hVEGF R3 cell lysate and analyzed using this DuoSet IC ELISA. Competition was observed only with recombinant human VEGF R3.

Product Datasheets

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Preparation and Storage

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: VEGFR3/Flt-4

VEGF R1 (Flt-1), VEGF R2 (KDR/Flk-1), and VEGF R3 (Flt-4) belong to the class III subfamily of receptor tyrosine kinases (RTKs). All three receptors contain seven immunoglobulin-like repeats in their extracellular domain and kinase insert domains in their intracellular region. They are best known for regulating VEGF family-mediated vasculogenesis, angiogenesis, and lymphangiogenesis. They are also mediators of neurotrophic activity and regulators of hematopoietic development. VEGF R2 is thought to be the primary inducer of VEGF-mediated blood vessel growth, while VEGF R3 plays a significant role in VEGF-C and VEGF-D-mediated lymphangiogenesis.

Long Name:
Vascular Endothelial Growth Factor Receptor 3
Entrez Gene IDs:
2324 (Human); 14257 (Mouse)
Alternate Names:
EC 2.7.10; EC; FLT4; Flt-4; fms-related tyrosine kinase 4; LMPH1A; PCLFLT41; soluble VEGFR3 variant 1; soluble VEGFR3 variant 2; soluble VEGFR3 variant 3; Tyrosine-protein kinase receptor FLT4; vascular endothelial growth factor receptor 3; VEGF R3; VEGFR3; VEGFR-3; VEGFR3Fms-like tyrosine kinase 4

Citations for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Decorin suppresses tumor lymphangiogenesis: A mechanism to curtail cancer progression
    Authors: Mondal, DK;Xie, C;Buraschi, S;Iozzo, RV;
    bioRxiv : the preprint server for biology
    Species: Human
    Sample Types: Cell Lysates
  2. Anti-Vascular Endothelial Growth Factor C Antibodies Efficiently Inhibit the Growth of Experimental Clear Cell Renal Cell Carcinomas
    Authors: A Dumond, C Montemagno, V Vial, R Grépin, G Pagès
    Cells, 2021-05-17;10(5):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Proteolytic activation defines distinct lymphangiogenic mechanisms for VEGFC and VEGFD
    J Clin Invest, 2016-05-09;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  4. The propeptides of VEGF-D determine heparin binding, receptor heterodimerization, and effects on tumor biology.
    Authors: Harris, Nicole C, Davydova, Natalia, Roufail, Sally, Paquet-Fifield, Sophie, Paavonen, Karri, Karnezis, Tara, Zhang, You-Fang, Sato, Teruhiko, Rothacker, Julie, Nice, Edouard, Stacker, Steven A, Achen, Marc G
    J Biol Chem, 2013-02-12;288(12):8176-86.
    Species: Human
    Sample Types: Cell Lysates
  5. Differential effects of the vascular endothelial growth factor receptor inhibitor PTK787/ZK222584 on tumor angiogenesis and tumor lymphangiogenesis.
    Authors: Schomber T, Zumsteg A, Strittmatter K, Crnic I, Antoniadis H, Littlewood-Evans A, Wood J, Christofori G
    Mol. Cancer Ther., 2009-01-01;8(1):55-63.
    Species: Transgenic Mouse
    Sample Types: Tissue Homogenates


  1. Which phosphorylated sites are recognized by this assay?

    • This assay utilizes an anti-phosphorylated tyrosine monoclonal detection antibody, and it recognizes all phosphorylated tyrosine residues.

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Reviews for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

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Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA
By Anonymous on 10/29/2023
Sample Tested: Breast cancer tissue,Serum and Plasma

Whether it's tissue specimens, blood samples, or supernatant, we can clearly see the differences and obtain reliable test results.

Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA
By Jenna Nguyen on 10/17/2019
Sample Tested: 7TD1 mouse hybridoma cell line