Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Canine VEGF Quantikine ELISA Kit Summary
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
Cell Culture Supernates
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 29.1 | 123 | 531 | 32.8 | 128 | 495 |
| Standard Deviation | 1.9 | 5 | 18.4 | 2.8 | 6.4 | 33 |
| CV% | 6.5 | 4.1 | 3.5 | 8.5 | 5 | 6.7 |
Serum, EDTA Plasma, Heparin Plasma
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 53.7 | 235 | 910 | 64.5 | 250 | 1003 |
| Standard Deviation | 3.6 | 10.6 | 46.2 | 5.7 | 17.4 | 61.7 |
| CV% | 6.7 | 4.5 | 5.1 | 8.8 | 7 | 6.2 |
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.
Scientific Data
Product Datasheets
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples and standards be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- 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. - 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. - Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process twice for a total of 3 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 3 times.
- Add 200 µL Substrate Solution to each well. PROTECT FROM LIGHT.
- For Cell Culture Supernate Samples: Incubate at room temperature for 20 minutes.
For Serum & Plasma Samples: Incubate at room temperature for 25 minutes. - 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.
50 µL Assay Diluent for Cell Culture Supernate Samples or 100 µL Assay Diluent for Serum/Plasma Samples
200 µL Standard or Sample for Cell Culture Supernate Samples or 100 µL Standard or Sample for Serum/Plasma Samples
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Citations for Canine VEGF Quantikine ELISA Kit
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.
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Citations: Showing 1 - 9
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Impairment of the Retinal Endothelial Cell Barrier Induced by Long-Term Treatment with VEGF-A165 No Longer Depends on the Growth Factor's Presence
Authors: HL Deissler, M Rehak, A Wolf
Biomolecules, 2022-05-23;12(5):.
Species: Bovine
Sample Types: Cell Culture Supernates
-
Association of markers of endothelial activation and dysfunction with occurrence and outcome of pulmonary hemorrhage in dogs with leptospirosis
Authors: F Sonderegge, A Nentwig, A Schweighau, T Francey, E Marti, J Mirkovitch, S Schuller
Journal of veterinary internal medicine, 2021-06-02;0(0):.
Species: Canine
Sample Types: Serum
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Angiogenic response in an in vitro model of dog microvascular endothelial cells stimulated with antigenic extracts from Dirofilaria immitis adult worms
Authors: T Zueva, R Morchón, E Carretón, JA Montoya-Al, A Santana, MD Bargues, S Mas-Coma, A Rodríguez-, F Simón
Parasit Vectors, 2019-06-24;12(1):315.
Species: Canine
Sample Types: Cell Culture Supernates
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Clinical Assessment of Intravenous Endothelial Progenitor Cell Transplantation in Dogs
Authors: SH Lee, JC Ra, HJ Oh, MJ Kim, EMN Setyawan, YB Choi, JW Yang, SK Kang, SH Han, GA Kim, BC Lee
Cell Transplant, 2019-04-24;0(0):9636897188216.
Species: Human
Sample Types: Serum
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In-vitro binding analysis of anti-human vascular endothelial growth factor antibodies bevacizumab and aflibercept with canine, feline, and equine vascular endothelial growth factor
Authors: LM Muellerlei, B Buxbaum, B Nell, DA Fux
Res. Vet. Sci., 2019-03-22;124(0):233-238.
Species: Canine
Sample Types: Recombinant Protein
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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., 2017-07-18;42(4):1493-1513.
Species: Bovine
Sample Types: Cell Culture Supernates
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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., 2017-04-07;0(0):.
Species: Feline
Sample Types: Plasma
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Pilot study to determine the feasibility of radiation therapy for dogs with right atrial masses and hemorrhagic pericardial effusion
Authors: MW Nolan, MM Arkans, D LaVine, T DeFrancesc, JA Myers, EH Griffith, LP Posner, BW Keene, SP Tou, TL Gieger
J Vet Cardiol, 2017-03-07;0(0):.
Species: Canine
Sample Types: Serum
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Outcomes of Spatially Fractionated Radiotherapy (GRID) for Bulky Soft Tissue Sarcomas in a Large Animal Model
Authors: MW Nolan, TL Gieger, AA Karakashia, MN Nikolova-K, LP Posner, DM Roback, JN Rivera, S Chang
Technol. Cancer Res. Treat, 2017-02-07;0(0):1533034617690.
Species: Canine
Sample Types: Serum
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