Home / VEGFR2/KDR/Flk-1 / Human sVEGFR2/KDR DuoSet ELISA

Human sVEGFR2/KDR DuoSet ELISA

Catalog #: DY357
8 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY357
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human VEGF R2 / KDR / Flk-1 ELISA Standard Curve
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Product Details
Procedure
Citations (8)
FAQs
Supplemental Products
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human sVEGFR2/KDR 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
31.2 - 2,000 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 VEGF R2/KDR. 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.

Normal Goat Serum: (Catalog # DY005)

 

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)

Normal Goat Serum: (Catalog # DY005)

 

Scientific Data

N/A Human VEGF R2 / KDR / Flk-1 ELISA Standard Curve View Larger

Human VEGF R2 / KDR / Flk-1 ELISA Standard Curve

Product Datasheets

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Product Datasheet
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SDS

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: VEGFR2/KDR/Flk-1

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 2
Entrez Gene IDs:
3791 (Human); 16542 (Mouse)
Alternate Names:
CD309 antigen; CD309; EC 2.7.10; EC 2.7.10.1; Fetal liver kinase 1; fetal liver kinase-1; Flk1; Flk-1; FLK1tyrosine kinase growth factor receptor; KDR; kinase insert domain receptor (a type III receptor tyrosine kinase); Kinase insert domain receptor; KRD1; Ly73; Protein-tyrosine kinase receptor flk-1; soluble VEGFR2; vascular endothelial growth factor receptor 2; VEGF R2; VEGFR; VEGFR2; VEGFR-2

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 with NGS, 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 sVEGFR2/KDR 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.

8 Citations: Showing 1 - 8
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  1. The Association of Biochemical and Genetic Biomarkers in VEGF Pathway with Depression
    Authors: FDD Nunes, LP Ferezin, SC Pereira, FV Figaro-Dru, LC Pinheiro, IC Menezes, CVW Baes, FB Coeli-Lacc, JE Tanus-Sant, MF Juruena, R Lacchini
    Pharmaceutics, 2022-12-09;14(12):.
    Species: Human
    Sample Types: Plasma
  2. Retinal VEGF-A Overexpression Is Not Sufficient to Induce Lymphangiogenesis Regardless of VEGF-C Upregulation and Lyve1+ Macrophage Infiltration
    Authors: I Wada, S Nakao, M Yamaguchi, Y Kaizu, M Arima, S Sawa, KH Sonoda
    Investigative Ophthalmology & Visual Science, 2021-10-04;62(13):17.
    Species: Human
    Sample Types: Vitreous Humor
  3. Hepatic blood flow by perfusion computed tomography as an imaging biomarker for patients with gastric cancer
    Authors: K Shuto, M Mori, C Kosugi, K Narushima, S Nakabayash, T Fujisiro, A Sato, K Hayano, H Shimizu, K Koda
    Oncol Lett, 2019-01-25;17(3):3267-3276.
    Species: Human
    Sample Types: Serum
  4. 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
  5. 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
  6. Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
    Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
    BMC Infect Dis, 2014-01-20;14(0):35.
    Species: Human
    Sample Types: Serum
  7. Differential expression of Vegfr-2 and its soluble form in preeclampsia.
    Authors: Munaut C, Lorquet S, Pequeux C, Coulon C, Le Goarant J, Chantraine F, Noel A, Goffin F, Tsatsaris V, Subtil D, Foidart JM
    PLoS ONE, 2012-03-12;7(3):e33475.
    Species: Human
    Sample Types: Plasma
  8. Expression of soluble vascular endothelial growth factor receptor-1 in human monocyte-derived mature dendritic cells contributes to their antiangiogenic property.
    Authors: Kishuku M, Nishioka Y, Abe S, Kishi J, Ogino H, Aono Y, Azuma M, Kinoshita K, Batmunkh R, Rentsenhand B, Makino H, Ranjan P, Minakuchi K, Sone S
    J. Immunol., 2009-12-15;183(12):8176-85.
    Species: Human
    Sample Types: Cell Culture Supernates

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