• Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (200 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
  • Sensitivity
    19.5 pg/mL
  • Assay Range
    19.5 - 1,250 pg/mL (Cell Culture Supernates), 39.1 - 2,500 pg/mL (Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant canine VEGF. This assay cross-reacts with Human VEGF165.
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity observed with 1 or more species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Canine VEGF Immunoassay is a 4.5 hour solid phase ELISA designed to measure VEGF levels in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant canine VEGF and antibodies raised against the recombinant protein. Results obtained for naturally occurring canine VEGF showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural canine VEGF.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean53.723591064.52501003
Standard Deviation3.610.646.25.717.461.7
CV%6.74.55.18.876.2

Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean29.112353132.8128495
Standard Deviation1.9518.42.86.433
CV%6.54.13.58.556.7

Recovery

The recovery of canine VEGF spiked to three different levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 102 94-110
EDTA Plasma (n=4) 98 92-107
Heparin Plasma (n=4) 107 96-118
Serum (n=4) 103 87-113
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of canine VEGF and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Canine VEGF Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: VEGF
VEGF (Vascular endothelial growth factor) is a potent mediator of both angiogenesis and vasculogenesis in the fetus and adult. It is required for regulating the proliferation, migration, and survival of embryonic endothelial cells and during wound healing and the female reproductive cycle in adults. Pathologically, it is involved in tumor development and tumor vascular leakage. VEGF binds to VEGF R1/Flt-1, VEGF R2/Flk-1/KDR, and Neuropilin-1. Multiple forms of VEGF are generated by alternative splicing and proteolysis.
    • Long Name
      Vascular Endothelial Growth Factor
    • Entrez Gene IDs
      7422 (Human); 22339 (Mouse); 83785 (Rat); 281572 (Bovine); 403802 (Canine); 493845 (Feline); 30682 (Zebrafish);
    • Alternate Names
      MVCD1; VAS; vascular endothelial growth factor A; Vascular permeability factor; Vasculotropin; VEGF; VEGFA; VEGF-A; VEGFMGC70609; VPF; VPFvascular endothelial growth factor;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples and standards be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent for Cell Culture Supernate Samples or 100 µL Assay Diluent for Serum/Plasma Samples
    4.   For Cell Culture Supernate Samples:  Add 50 µL of Assay Diluent to each well.
      For Serum & Plasma Samples:  Add 100 µL of Assay Diluent to each well.

    5. 200 µL Standard or Sample for Cell Culture Supernate Samples or 100 µL Standard or Sample for Serum/Plasma Samples
    6.   For Cell Culture Supernate Samples:  Add 200 µL of Standard or sample to each well.
      For Serum & Plasma Samples:  Add 100 µL of Standard or sample to each well.
    7.   Cover with a plate sealer, and incubate at room temperature for 2 hours.
    8.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

    9. 200 µL Conjugate
    10.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    11.   Aspirate and wash 3 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. PROTECT FROM LIGHT.
    14. For Cell Culture Supernate Samples:  Incubate at room temperature for 20 minutes.
      For Serum & Plasma Samples:  Incubate at room temperature for 25 minutes.

    15. 50 µL Stop Solution
    16. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    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.

    2 Citations: Showing 1 - 2
    Filter your results:

    Species
    Sample Type
    1. Plasma N-Terminal Probrain Natriuretic Peptide, Vascular Endothelial Growth Factor, and Cardiac Troponin I as Novel Biomarkers of Hypertensive Disease and Target Organ Damage in Cats
      Authors: ES Bijsmans, RE Jepson, C Wheeler, HM Syme, J Elliott
      J. Vet. Intern. Med., 2017;0(0):.
      Species: Feline
      Sample Type: Plasma
    2. Inhibition of Single Routes of Intracellular Signaling is Not Sufficient to Neutralize the Biphasic Disturbance of a Retinal Endothelial Cell Barrier Induced by VEGF-A165
      Authors: HL Deissler, GK Lang, GE Lang
      Cell. Physiol. Biochem., 2017;42(4):1493-1513.
      Species: Bovine
      Sample Type: Cell Culture Supernates
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