Human VEGF Quantikine QuicKit ELISA
R&D Systems, part of Bio-Techne | Catalog # QK293
Key Product Details
Assay Length
80 minutes
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Saliva (25 µL), Urine (13 µL), Human Milk (10 µL)
Sensitivity
3.19 pg/mL
Assay Range
28.1-1800 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
Product Summary for Human VEGF Quantikine QuicKit ELISA
The Quantikine™ QuicKit™ Human VEGF Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human VEGF levels in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains Sf 21-expressed recombinant human VEGF and antibodies raised against the recombinant protein. Results obtained using natural human VEGF showed linear curves that were parallel to the standard curves obtained using the QuicKit standards. These results indicate that this kit can be used to determine relative mass values for natural human VEGF.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Natural and recombinant human VEGF.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma/Saliva/Urine/Human Milk - Samples from apparently healthy volunteers were
evaluated for the presence of human VEGF in this assay. No medical histories were available for
the donors used in this study.
Cell Culture Supernates:
| Sample Type | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=10) | 519 | 175-1093 | 352 |
| EDTA plasma (n=10) | 79.4 | 40.6-183 | 43.1 |
| Heparin plasma (n=10) | 69.8 | 38.9-105 | 23.7 |
| Saliva (n=4) | 3521 | 2055-4880 | 1374 |
| Urine (n=4) | 566 | 219-1184 | 429 |
| Human milk (n=4) | 27,535 | 23,100-31,888 | 4620 |
Cell Culture Supernates:
PBMCs were isolated from a single donor over a Ficoll-Paque PLUS density gradient. PBMCs
were seeded at 1 x 106
/mL and cultured in RPMI 1640 supplemented with 10% fetal bovine
serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells
were stimulated with 10 μg/mL PHA for 5 days. Aliquots of the cell culture supernatants were
removed, assayed for human VEGF, and measured 169 pg/mL.
JEG-3 human epithelial choriocarcinoma cells were cultured in MEM Earle's Salts and
supplemented with 10% FBS. Cells were stimulated with 100 ng/mL of LPS for 24 hours. An
aliquot of the cell culture supernate was removed, assayed for human VEGF, and measured
11,100 pg/mL.
THP-1 acute monocytic leukemia cells were cultured in RPMI and supplemented with 10% fetal
bovine serum. Cells were stimulated with 1 μg/mL Retionic Acid and 100 ng/mL PMA for 4 days.
An aliquot of the cell culture supernate was removed, assayed for human VEGF, and measured
1829 pg/mL.
PC-3 human prostate cancer cells were cultured in RPMI and supplemented with 10% fetal
bovine serum until confluent. An aliquot of the cell culture supernate was removed, assayed for
human VEGF, and measured 2366 pg/mL.
Precision
Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Human Milk, Saliva, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||
|---|---|---|---|---|
| Sample | 1 | 2 | 1 | 2 |
| n | 20 | 20 | 10 | 10 |
| Mean (pg/mL) | 209 | 1193 | 203 | 1216 |
| Standard Deviation | 7.45 | 35.1 | 18.3 | 56.5 |
| CV% | 3.6 | 2.9 | 9.0 | 4.6 |
Recovery for Human VEGF Quantikine QuicKit ELISA
The recovery of human VEGF spiked to three levels in samples throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 99 | 94-104 |
| EDTA Plasma (n=2) | 83 | 79-88 |
| Heparin Plasma (n=2) | 77 | 70-81 |
| Human Milk (n=2) | 102 | 98-105 |
| Saliva (n=2) | 104 | 97-115 |
| Serum (n=2) | 79 | 74-84 |
| Urine (n=2) | 78 | 70-85 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human VEGF were diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human VEGF Quantikine QuicKit ELISA
Human VEGF Standard Curve
Human VEGF QuicKit ELISA Spiked Linearity Competitor Comparison
Samples containing VEGF were diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 99%-107% compared to 100%-145% for the top competitor.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: VEGF
VEGF dimers bind to two related receptor tyrosine kinases, VEGF R1 (also called Flt-1) and VEGF R2 (Flk-1/KDR), and induce their homodimerization and autophosphorylation (3, 4, 7, 17, 18). These receptors have seven extracellular immunoglobulin-like domains and an intracellular split tyrosine kinase domain. They are expressed on vascular endothelial cells and a range of non-endothelial cells. Although VEGF affinity is highest for binding to VEGF R1, VEGF R2 appears to be the primary mediator of VEGF angiogenic activity (3, 4). VEGF165 also binds the semaphorin receptor, neuropilin-1, which promotes complex formation with VEGF R2 (19).
VEGF is best known for its role in vasculogenesis. During embryogenesis, VEGF regulates the proliferation, migration, and survival of endothelial cells (3, 4), thus regulating blood vessel density and size, but playing no role in determining vascular patterns. VEGF promotes bone formation through osteoblast and chondroblast recruitment and is also a monocyte chemoattractant (20-22). After birth, VEGF maintains endothelial cell integrity and is a potent mitogen for micro- and macro-vascular endothelial cells. In adults, VEGF functions mainly in wound healing and the female reproductive cycle (3). In diseased tissues, VEGF promotes vascular permeability. It is thus thought to contribute to tumor metastasis by promoting both extravasation and tumor angiogenesis (23, 24). Various strategies have been employed therapeutically to antagonize VEGF-mediated tumor angiogenesis (25). Circulating VEGF levels correlate with disease activity in autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and systemic lupus erythematosus (26).
Long Name
Vascular Endothelial Growth Factor
Alternate Names
MVCD1, VAS, Vasculotropin, VEGF-A, VEGFA, VPF
Entrez Gene IDs
Gene Symbol
VEGFA
Additional VEGF Products
Product Documents for Human VEGF Quantikine QuicKit ELISA
Product Specific Notices for Human VEGF Quantikine QuicKit ELISA
For research use only
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