Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
Complete Your Research
Complete Your Experiment
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
Preparation and Storage
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.
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.
4
Citations: Showing 1 - 4
Filter your results:
Filter by:
-
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;10(5):.
Species: Human
Sample Types: Cell Culture Supernates
-
Proteolytic activation defines distinct lymphangiogenic mechanisms for VEGFC and VEGFD
J Clin Invest, 2016;0(0):.
Species: Human
Sample Types: Cell Lysates
-
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;288(12):8176-86.
Species: Human
Sample Types: Cell Lysates
-
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;8(1):55-63.
Species: Transgenic Mouse
Sample Types: Tissue Homogenates
FAQs
-
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.
DuoSet Ancillary Reagent Kits
Supplemental ELISA Products
Reviews for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA
There are currently no reviews for this product. Be the first to review Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA and earn rewards!
Have you used Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image