Human VEGF QuantiGlo ELISA Kit

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QVE00B
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Human VEGF Chemiluminescent ELISA Cell Culture Supernate/Serum/Plasma/Saliva Standard Curve
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Human VEGF QuantiGlo ELISA Kit Summary

Assay Length
5.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Saliva (50 uL), Urine (50 uL)
Sensitivity
5.99 pg/mL
Assay Range
6.4 - 20,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
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.< 50% cross-species reactivity observed with species tested.
Interference
Interference observed with 1 or more available related molecules.

Product Summary

The QuantiGlo Human VEGF Chemiluminescent Immunoassay is a 5.5 hour solid phase ELISA designed to measure VEGF165 levels in cell culture supernates, serum, plasma, saliva, and urine. 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 QuantiGlo kit standards. These results indicate that this kit can be used to determine relative mass values for natural human VEGF.

Precision

Intra-Assay Precision (Precision within an assay) Four samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Four samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 4 1 2 3 4
n 20 20 20 20 20 20 20 20
Mean (pg/mL) 22.2 392 1857 13972 22.7 389 1843 13411
Standard Deviation 1.76 12.9 52.3 557 2 27.6 131 562
CV% 7.9 3.3 2.8 4 8.8 7.1 7.1 4.2

Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 4 1 2 3 4
n 20 20 20 20 20 20 20 20
Mean (pg/mL) 24.4 417 1999 14245 22.9 383 1823 12825
Standard Deviation 1.24 16 44.9 608 2.36 28.9 141 536
CV% 5.1 3.8 2.2 4.3 10.3 7.5 7.7 4.2

Recovery

The recovery of 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=4) 103 95-113
EDTA Plasma (n=4) 104 98-110
Heparin Plasma (n=4) 104 96-111
Serum (n=4) 102 90-112
Urine (n=4) 99 90-106

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of VEGF in various matrices were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human VEGF Chemiluminescent ELISA Linearity

Scientific Data

Human VEGF Chemiluminescent ELISA Cell Culture Supernate/Serum/Plasma/Saliva Standard Curve

Human VEGF Chemiluminescent ELISA Urine Standard Curve

Product Datasheets

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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

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
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

Citations for Human VEGF QuantiGlo 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.

23 Citations: Showing 1 - 10
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  1. Development of a potency assay for CD34+ cell-based therapy
    Authors: Aries, A;Vignon, C;Zanetti, C;Goubaud, A;Cormier, A;Diederichs, A;Lahlil, R;Hénon, P;Garitaonandia, I;
    Scientific reports
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Regenerating glomerular metabolism and function by podocyte pyruvate kinase M2 in diabetic nephropathy
    Authors: J Fu, T Shinjo, Q Li, R St-Louis, K Park, MG Yu, H Yokomizo, F Simao, Q Huang, IH Wu, GL King
    JCI Insight, 2022-03-08;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  3. Downregulation of tumor suppressive microRNAs in vivo in dense breast tissue of postmenopausal women
    Authors: A Abrahamsso, A Capodanno, A Rzepecka, C Dabrosin
    Oncotarget, 2017-09-15;8(54):92134-92142.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. MicroRNA-1 acts as a tumor suppressor microRNA by inhibiting angiogenesis-related growth factors in human gastric cancer
    Authors: M Xie, DA Dart, T Guo, XF Xing, XJ Cheng, H Du, WG Jiang, XZ Wen, JF Ji
    Gastric Cancer, 2017-05-10;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Hypoxic Preconditioning Increases Survival and Pro-Angiogenic Capacity of Human Cord Blood Mesenchymal Stromal Cells In Vitro.
    Authors: Bader A, Klose K, Bieback K, Korinth D, Schneider M, Seifert M, Choi Y, Kurtz A, Falk V, Stamm C
    PLoS ONE, 2015-09-18;10(9):e0138477.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Aging impairs VEGF-mediated, androgen-dependent regulation of angiogenesis.
    Authors: Lecce, Laura, Lam, Yuen Tin, Lindsay, Laura A, Yuen, Sui Chin, Simpson, Philippa, Handelsman, David J, Ng, Martin K
    Mol Endocrinol, 2014-07-24;28(9):1487-501.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Inflammation induced by MMP-9 enhances tumor regression of experimental breast cancer.
    Authors: Leifler K, Svensson S, Abrahamsson A, Bendrik C, Robertson J, Gauldie J, Olsson A, Dabrosin C
    J Immunol, 2013-03-15;190(8):4420-30.
    Species: Mouse
    Sample Types: Tissue Homogenates
  8. Angiogenic and vasculogenic factors in the vitreous from patients with proliferative diabetic retinopathy.
    Authors: Abu El-Asrar, Ahmed M, Nawaz, Mohd Imt, Kangave, Dustan, Mairaj Siddiquei, Mohammed, Geboes, Karel
    J Diabetes Res, 2013-03-10;0(0):539658.
    Species: Human
    Sample Types: Cell Lysates
  9. Vascular endothelial growth factor-A (VEGF-A) and chemokine ligand-2 (CCL2) in Amyotrophic Lateral Sclerosis (ALS) patients.
    Authors: Gupta PK, Prabhakar S
    J Neuroinflammation, 2011-05-13;8(1):47.
    Species: Human
    Sample Types: CSF
  10. Overexpression of tumor vascular endothelial growth factor A may portend an increased likelihood of progression in a phase II trial of bevacizumab and erlotinib in resistant ovarian cancer.
    Authors: Chambers SK, Clouser MC, Baker AF, Roe DJ, Cui H, Brewer MA, Hatch KD, Gordon MS, Janicek MF, Isaacs JD, Gordon AN, Nagle RB, Wright HM, Cohen JL, Alberts DS
    Clin. Cancer Res., 2010-11-01;16(21):5320-8.
    Species: Human
    Sample Types: Urine
  11. Dietary lariciresinol attenuates mammary tumor growth and reduces blood vessel density in human MCF-7 breast cancer xenografts and carcinogen-induced mammary tumors in rats.
    Authors: Saarinen NM, Warri A, Dings RP, Airio M, Smeds AI, Mäkelä S
    Int. J. Cancer, 2008-09-01;123(5):1196-204.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Gene transfer of matrix metalloproteinase-9 induces tumor regression of breast cancer in vivo.
    Authors: Bendrik C, Robertson J, Gauldie J, Dabrosin C
    Cancer Res., 2008-05-01;68(9):3405-12.
    Species: Human
    Sample Types: Tissue Homogenates
  13. Clinical correlations and prognostic relevance of HGF, VEGF AND FGF expression in Brazilian patients with non-Hodgkin lymphoma.
    Authors: Etto L, Lacerda E, Baiocchi O, Silva V, Dalboni M, Alves A, Silva M, Vettore A, Colleoni G
    Leuk. Lymphoma, 2008-02-01;49(2):257-64.
    Species: Human
    Sample Types: Serum
  14. Regulation of angiogenic factors by HDM2 in renal cell carcinoma.
    Authors: Carroll VA, Ashcroft M
    Cancer Res., 2008-01-15;68(2):545-52.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Sphingosine kinase 1 is up-regulated during hypoxia in U87MG glioma cells. Role of hypoxia-inducible factors 1 and 2.
    Authors: Anelli V, Gault CR, Cheng AB, Obeid LM
    J. Biol. Chem., 2007-11-30;283(6):3365-75.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Where is VEGF in the body? A meta-analysis of VEGF distribution in cancer.
    Authors: Kut C, Mac Gabhann F, Popel AS
    Br. J. Cancer, 2007-10-02;97(7):978-85.
    Species: Human
    Sample Types: Serum
  17. Very low density lipoprotein receptor, a negative regulator of the wnt signaling pathway and choroidal neovascularization.
    Authors: Chen Y, Hu Y, Lu K, Flannery JG, Ma JX
    J. Biol. Chem., 2007-09-23;282(47):34420-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Selective inhibition of MEK1/2 reveals a differential requirement for ERK1/2 signalling in the regulation of HIF-1 in response to hypoxia and IGF-1.
    Authors: Sutton KM, Hayat S, Chau NM, Cook S, Pouyssegur J, Ahmed A, Perusinghe N, Le Floch R, Yang J, Ashcroft M
    Oncogene, 2007-01-08;26(27):3920-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Role of hypoxia-inducible factor (HIF)-1alpha versus HIF-2alpha in the regulation of HIF target genes in response to hypoxia, insulin-like growth factor-I, or loss of von Hippel-Lindau function: implications for targeting the HIF pathway.
    Authors: Carroll VA, Ashcroft M
    Cancer Res., 2006-06-15;66(12):6264-70.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Dynamic contrast-enhanced MRI to quantify VEGF-enhanced tissue-engineered bladder graft neovascularization: pilot study.
    Authors: Cartwright L, Farhat WA, Sherman C, Chen J, Babyn P, Yeger H, Cheng HL
    J Biomed Mater Res A, 2006-05-01;77(2):390-5.
    Species: Rabbit
    Sample Types: Cell Culture Supernates
  21. Protease-activated receptor-2 regulates vascular endothelial growth factor expression in MDA-MB-231 cells via MAPK pathways.
    Authors: Mueller BM
    Biochem. Biophys. Res. Commun., 2006-04-19;344(4):1263-70.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Increased vascular endothelial growth factor and receptors: relationship to angiogenesis in asthma.
    Authors: Feltis BN, Wignarajah D, Zheng L, Ward C, Reid D, Harding R, Walters EH
    Am. J. Respir. Crit. Care Med., 2006-03-09;173(11):1201-7.
    Species: Human
    Sample Types: BALF
  23. Evaluating hypoxia-inducible factor-1alpha as a cancer therapeutic target via inducible RNA interference in vivo.
    Authors: Li L, Lin X, Staver M, Shoemaker A, Semizarov D, Fesik SW, Shen Y
    Cancer Res., 2005-08-15;65(16):7249-58.
    Species: Human
    Sample Types: Tissue Homogenates

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