Natural and recombinant canine VEGF. This assay cross-reacts with Human VEGF165.
Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity observed with 1 or more species tested.
Interference observed with 1 or more available related molecules.
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.
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
Cell Culture Supernates
The recovery of canine VEGF spiked to three different levels throughout the range of the assay was evaluated.
Average % Recovery
Cell Culture Media (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
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.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
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.
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.
50 µL Assay Diluent for Cell Culture Supernate Samples or 100 µL Assay Diluent for Serum/Plasma Samples
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.
200 µL Standard or Sample for Cell Culture Supernate Samples or 100 µL Standard or Sample for Serum/Plasma Samples
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.
200 µL Conjugate
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.
200 µL Substrate Solution
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.
50 µL Stop Solution
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.
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