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Human sVEGFR3/Flt-4 DuoSet ELISA

Catalog #: DY349
10 Citations 1 Review Datasheet / COA / SDS
Newer Version Available: DY349B-05
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DY349
Human VEGF R3 / Flt-4 ELISA Standard Curve
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Product Details
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Citations (10)
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Reviews (1)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human sVEGFR3/Flt-4 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
100 µL
Assay Range
156.0 - 10,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
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 ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human sVEGF R3/Flt-4. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

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)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A Human VEGF R3 / Flt-4 ELISA Standard Curve View Larger

Human VEGF R3 / Flt-4 ELISA Standard Curve

Product Datasheets

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Product Datasheet
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Preparation and Storage

Shipping
The product is shipped at ambient temperature. 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 2.7.10.1; 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human sVEGFR3/Flt-4 DuoSet 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.

10 Citations: Showing 1 - 10
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  1. Tuberculosis-diabetes co-morbidity is characterized by heightened systemic levels of circulating angiogenic factors.
    Authors: Kumar N, Moideen K, Sivakumar S, Menon P, Viswanathan V, Kornfeld H, Babu S
    J Infect, 2016-10-04;74(1):10-21.
    Species: Human
    Sample Types: Plasma
  2. Circulating Angiogenic Factors as Biomarkers of Disease Severity and Bacterial Burden in Pulmonary Tuberculosis.
    Authors: Kumar N, Banurekha V, Nair D, Babu S
    PLoS ONE, 2016-01-04;11(1):e0146318.
    Species: Human
    Sample Types: Plasma
  3. Cytomegalovirus impairs cytotrophoblast-induced lymphangiogenesis and vascular remodeling in an in vivo human placentation model.
    Authors: Tabata T, Petitt M, Fang-Hoover J, Rivera J, Nozawa N, Shiboski S, Inoue N, Pereira L
    Am J Pathol, 2012-09-07;181(5):1540-59.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Plasma biomarkers as predictors of outcome in patients with advanced hepatocellular carcinoma.
    Authors: Llovet JM, Pena CE, Lathia CD, Shan M, Meinhardt G, Bruix J, Pallota MG, Zarba JJ, Boyer M, Riordan S, Strickland A, Tebbutt N, Thomson B, Borbath I, De Greve J, Van Laethem J-, Van Steenbergen W, Van Vlierberghe H, Barrios C, Cosme de Oliveira A, Kotzev I, Takov D, Tchernev K, Burak K, Ma M, Metrakos P, Olweny C, Sherman M, Gamargo Garate C, Martinez-Castillo J, Beaugrand M, Bennouna J, Blanc J-, Bronowicki J-, Degos F, Dominguez S, Grange J-, Hillon P, Raoul J-, Seitz J-, Blum H, Buggisch P, Caspary W, Dollinger M, Gerken G, Goke B, Gregor M, Greten T, Haussinger D, Hilgard P, Scherübl J, Scheulen M, Schmid R, Spengler U, Wiest R, Zeuzem S, Arvanitakis C, Germanidis G, Katsos I, Figer A, Stemmer S, Amadori D, Bolondi L, Cognetti F, Craxi A, Farinati F, Gridelli C, Martoni A, Mazzaferro V, Porta C, Ricci S, Sangiovanni A, Santoro A, Trevisani F, Cisnero Garza LE, Gane E, O'Donnell A, Leon J, Lozano A, Jassem J, Rydzewska G, Szawlowski A, Tomczak P, Badulescu F, Miron L, Kubyshkin V, Bruix J, Forner A, Bustamante Schneider J, Diago M, Montero Alvarez JL, Pascual S, Ruíz del Arbol L, Sangro B, Solá R, Tabernero J, Muellhaupt B, Roth A, Jeffry Evans TR, Falk S, Meyer T, Reeves H, Ross P, Befeler A, Boyer T, Britten C, Byrne T, Garcia-Tsao G, Gold P, Goldenberg A, Heuman D, Kennedy P, Koch A, Llovet JM, Marrero J, Schilsky M, Schwartz J, Schwartz M
    Clin. Cancer Res., 2012-02-28;18(8):2290-300.
    Species: Human
    Sample Types: Plasma
  5. Mechanism-related circulating proteins as biomarkers for clinical outcome in patients with unresectable hepatocellular carcinoma receiving sunitinib.
    Authors: Harmon CS, DePrimo SE, Raymond E, Cheng AL, Boucher E, Douillard JY, Lim HY, Kim JS, Lechuga MJ, Lanzalone S, Lin X, Faivre S
    J Transl Med, 2011-07-25;9(0):120.
    Species: Human
    Sample Types: Plasma
  6. Antiangiogenic gene therapy with soluble VEGFR-1, -2, and -3 reduces the growth of solid human ovarian carcinoma in mice.
    Authors: Sallinen H, Anttila M, Narvainen J, Koponen J, Hamalainen K, Kholova I, Heikura T, Toivanen P, Kosma VM, Heinonen S, Alitalo K, Yla-Herttuala S
    Mol. Ther., 2008-12-02;17(2):278-84.
    Species: Human
    Sample Types: Plasma
  7. Antitumor activity and biomarker analysis of sunitinib in patients with bevacizumab-refractory metastatic renal cell carcinoma.
    Authors: Rini BI, Michaelson MD, Rosenberg JE, Bukowski RM, Sosman JA, Stadler WM, Hutson TE, Margolin K, Harmon CS, DePrimo SE, Kim ST, Chen I, George DJ
    J. Clin. Oncol., 2008-08-01;26(22):3743-8.
    Species: Human
    Sample Types: Plasma
  8. Phase II study of sunitinib malate, an oral multitargeted tyrosine kinase inhibitor, in patients with metastatic breast cancer previously treated with an anthracycline and a taxane.
    Authors: Burstein HJ, Elias AD, Rugo HS, Cobleigh MA, Wolff AC, Eisenberg PD, Lehman M, Adams BJ, Bello CL, DePrimo SE, Baum CM, Miller KD
    J. Clin. Oncol., 2008-03-17;26(11):1810-6.
    Species: Human
    Sample Types: Plasma
  9. Single-nucleotide polymorphisms and haplotypes in the VEGF receptor 3 gene and the haplotype GC in the VEGFA gene are associated with psoriasis in Koreans.
    Authors: Lee JH, Cho EY, Namkung JH, Kim E, Kim S, Shin ES, Lee JE, Yang JM
    J. Invest. Dermatol., 2007-12-20;128(6):1599-603.
    Species: Human
    Sample Types: Serum
  10. Phase II trial of sunitinib in patients with metastatic colorectal cancer after failure of standard therapy.
    Authors: Saltz LB, Rosen LS, Marshall JL, Belt RJ, Hurwitz HI, Eckhardt SG, Bergsland EK, Haller DG, Lockhart AC, Rocha Lima CM, Huang X, DePrimo SE, Chow-Maneval E, Chao RC, Lenz HJ
    J. Clin. Oncol., 2007-10-20;25(30):4793-9.
    Species: Human
    Sample Types: Plasma

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Human VEGFR3/Flt-4 DuoSet ELISA
By Jenna Nguyen on 10/17/2019
Sample Tested: 4T1 mouse breast cancer cell line