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Human VEGF ELISA Kit - Quantikine Best Seller

Bio-Techne includes R&D Systems | Catalog # DVE00

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DVE00
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SVE00

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (200 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Citrate Plasma (100 µL)

Sensitivity

5 pg/mL (Cell Culture Supernates)
9 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)

Assay Range

15.6-1000 pg/mL (Cell Culture Supernates)
31.3-2000 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)

Product Summary for Human VEGF Quantikine ELISA Kit

The Quantikine Human VEGF Immunoassay is a 4.5 hour solid phase ELISA designed to measure VEGF165 levels in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human VEGF165 and antibodies raised against the recombinant protein. Results obtained for naturally occurring human VEGF and recombinant human VEGF121 showed linear curves that were parallel to the standard curves obtained using the Quantikine Human VEGF Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human VEGF.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human

Specificity

Natural and recombinant human VEGF. This assay also recognizes recombinant human VEGF165b.

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.

Sample Values

Serum/Plasma - 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.

Sample TypeMean of detectable (pg/mL)% DetectableRange (pg/mL)
Serum (n=37)22010062-707
EDTA plasma (n=37)6124ND-115
Heparin plasma (n=37)4122ND-55
Citrate plasma (n=37)___0ND
ND=Non-Detectable

Cell Culture Supernates: 
Human peripheral blood mononuclear cells (10 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 5 days. Aliquots of the cell culture supernates were removed, assayed for levels of human VEGF, and measured 87.1 pg/mL and 1041 pg/mL, respectively.

PC-3 human prostate cancer cells were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 60 nM of PMA for 24 hours. Aliquots of the cell culture supernates were removed, assayed for levels of human VEGF, and measured 3373 pg/mL and 3652 pg/mL, respectively.

JEG-3 human epithelial choriocarcinoma cells were cultured in MEM + NEAA supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 100 ng/mL of LPS for 1 day. Aliquots of the cell culture supernates were removed, assayed for levels of human VEGF, and measured 1538 pg/mL and 1938 pg/mL, respectively. 

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.

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 5.0 18.4 2.8 6.4 33.0
CV% 6.5 4.1 3.5 8.5 5.0 6.7

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

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 7.0 6.2

Recovery for Human VEGF Quantikine ELISA Kit

The recovery of human VEGF spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 102 95-111
Citrate Plasma (n=5) 100 88-113
EDTA Plasma (n=5) 97 82-113
Heparin Plasma (n=5) 93 82-102
Serum (n=5) 102 92-115

Linearity

To assess linearity of the assay, samples were spiked with high concentrations of human VEGF and diluted with the appropriate calibrator diluent to produce samples with values within the dynamic range of the assay.

Human VEGF ELISA Linearity

Scientific Data Images for Human VEGF Quantikine ELISA Kit

Human VEGF ELISA Cell Culture Supernate Standard Curve

Human VEGF ELISA Cell Culture Supernate Standard Curve

Human VEGF ELISA Serum/Plasma Standard Curve

Human VEGF ELISA Serum/Plasma Standard Curve

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

Vascular endothelial growth factor (VEGF or VEGF-A), also known as vascular permeability factor (VPF), is a potent mediator of both angiogenesis and vasculogenesis in the fetus and adult (1-3). It is a member of the PDGF family that is characterized by the presence of eight conserved cysteine residues in a cystine knot structure and the formation of antiparallel disulfide-linked dimers (4). Humans express alternately spliced isoforms of 121, 145, 165, 183, 189, and 206 amino acids (aa) in length (4). VEGF165 appears to be the most abundant and potent isoform, followed by VEGF121 and VEGF189 (3, 4). Isoforms other than VEGF121 contain basic heparin-binding regions and are not freely diffusible (4). Human VEGF165 shares 88% aa sequence identity with corresponding regions of mouse and rat VEGF. VEGF is expressed in multiple cells and tissues including skeletal and cardiac muscle (5, 6), hepatocytes (7), osteoblasts (8), neutrophils (9), macrophages (10), keratinocytes (11), brown adipose tissue (12), CD34+ stem cells (13), endothelial cells (14), fibroblasts, and vascular smooth muscle cells (15). VEGF expression is induced by hypoxia and cytokines such as IL-1, IL-6, IL-8, oncostatin M, and TNF-alpha (3, 4, 9, 16). VEGF isoforms are differentially expressed during development and in the adult (3). 
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

7422 (Human); 22339 (Mouse); 83785 (Rat); 281572 (Bovine); 403802 (Canine); 493845 (Feline); 30682 (Zebrafish)

Gene Symbol

VEGFA

Additional VEGF Products

Product Documents for Human VEGF Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human VEGF Quantikine ELISA Kit

For research use only

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⚠ 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.