Mouse VEGF DuoSet ELISA

Catalog # Availability Size / Price Qty
DY493
DY493-05
Ancillary Products Available
Mouse VEGF ELISA Standard Curve
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Product Details
Procedure
Citations (33)
FAQs
Supplemental Products
Reviews (2)

Mouse VEGF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For five or fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse VEGF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Data Example

Mouse VEGF ELISA Standard Curve

Product Datasheets

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 in adults (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 anti-parallel disulfide-linked dimers (4). Alternately spliced isoforms of 120, 164 and 188 amino acids (aa) have been found in mice, while 121, 145, 165, 183, 189, and 206 aa isoforms have been identified in humans (2, 4). In humans, VEGF165 appears to be the most abundant and potent isoform, followed by VEGF121 and VEGF189 (3, 4). The same pattern may exist in mice. Isoforms other than VEGF120 and VEGF121 contain basic heparin-binding regions and are not freely diffusible (4). Mouse VEGF164 shares 97% aa sequence identity with corresponding regions of rat VEGF. It also shares 89% aa sequence identity with human and porcine VEGF, 88% with bovine VEGF, and 90% with feline, equine, and canine 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). The 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). In postnatal life, 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Mouse VEGF DuoSet ELISA

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.

33 Citations: Showing 1 - 10
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  1. Protein Kinase A Catalytic Subunit Is a Molecular Switch that Promotes the Pro-tumoral Function of Macrophages
    Authors: YR Na, JW Kwon, DY Kim, H Chung, J Song, D Jung, H Quan, D Kim, JS Kim, YW Ju, W Han, HS Ryu, YS Lee, JJ Hong, SH Seok
    Cell Rep, 2020;31(6):107643.
    Species: Mouse
    Sample Types:
  2. Functional Evaluation of AMD-Associated Risk Variants of Complement Factor B
    Authors: C Pilotti, J Greenwood, SE Moss
    Invest. Ophthalmol. Vis. Sci., 2020;61(5):19.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. Inhibition of FLT1 ameliorates muscular dystrophy phenotype by increased vasculature in a mouse model of Duchenne muscular dystrophy
    Authors: M Verma, Y Shimizu-Mo, Y Asakura, JP Ennen, J Bosco, Z Zhou, GH Fong, S Josiah, D Keefe, A Asakura
    PLoS Genet., 2019;15(12):e1008468.
    Species: Mouse
    Sample Types: Cell Culture Su
  4. A Virus-Derived Immune Modulating Serpin Accelerates Wound Closure with Improved Collagen Remodeling
    Authors: L Zhang, JR Yaron, AM Tafoya, SE Wallace, J Kilbourne, S Haydel, K Rege, G McFadden, AR Lucas
    J Clin Med, 2019;8(10):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  5. SOCS3 deficiency in myeloid cells promotes retinal degeneration and angiogenesis through arginase-1 up-regulation in experimental autoimmune uveoretinitis
    Authors: M Chen, J Zhao, IH Ali, S Marry, J Augustine, M Bhuckory, A Lynch, A Kissenpfen, H Xu
    Am. J. Pathol., 2018;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. Muscle Satellite Cell Cross-Talk with a Vascular Niche Maintains Quiescence via VEGF and Notch Signaling
    Authors: M Verma, Y Asakura, BSR Murakonda, T Pengo, C Latroche, B Chazaud, LK McLoon, A Asakura
    Cell Stem Cell, 2018;23(4):530-543.e9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. MCPIP1 downregulation in clear cell renal cell carcinoma promotes vascularization and metastatic progression
    Authors: P Marona, J Górka, Z Mazurek, W Wilk, J Rys, M Majka, J Jura, K Miekus
    Cancer Res., 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. Proangiogenic alginate-g-pyrrole hydrogel with decoupled control of mechanical rigidity and electrically conductivity
    Authors: RJ DeVolder, Y Seo, H Kong
    Biomater Res, 2017;21(0):24.
    Species: Mouse
    Sample Types: Whole Cells
  9. ECM-based macroporous sponges release essential factors to support the growth of hematopoietic cells
    J Control Release, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Interactions among Lung Cancer Cells, Fibroblasts, and Macrophages in 3D Co-Cultures and the Impact on MMP-1 and VEGF Expression
    Authors: Xiao-Qing Liu
    PLoS ONE, 2016;11(5):e0156268.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Tumor Microenvironment Remodeling by 4-Methylumbelliferone Boosts the Antitumor Effect of Combined Immunotherapy in Murine Colorectal Carcinoma.
    Authors: Malvicini M, Fiore E, Ghiaccio V, Piccioni F, Rizzo M, Olmedo Bonadeo L, Garcia M, Rodriguez M, Bayo J, Peixoto E, Atorrasagasti C, Alaniz L, Aquino J, Matar P, Mazzolini G
    Mol Ther, 2015;23(9):1444-55.
    Species: Mouse
    Sample Types: Serum
  12. VEGF-A modulates expression of inhibitory checkpoints on CD8+ T cells in tumors.
    Authors: Voron T, Colussi O, Marcheteau E, Pernot S, Nizard M, Pointet A, Latreche S, Bergaya S, Benhamouda N, Tanchot C, Stockmann C, Combe P, Berger A, Zinzindohoue F, Yagita H, Tartour E, Taieb J, Terme M
    J Exp Med, 2015;212(2):139-48.
    Species: Mouse
    Sample Types: Tissue Homogenates
  13. Cessation of CCL2 inhibition accelerates breast cancer metastasis by promoting angiogenesis.
    Authors: Bonapace L, Coissieux M, Wyckoff J, Mertz K, Varga Z, Junt T, Bentires-Alj M
    Nature, 2015;515(7525):130-3.
    Species: Mouse
    Sample Types: Tissue Homogenates
  14. The monocyte to macrophage transition in the murine sterile wound.
    Authors: Crane, Meredith, Daley, Jean M, van Houtte, Olivier, Brancato, Samielle, Henry, William, Albina, Jorge E
    PLoS ONE, 2014;9(1):e86660.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  15. Advances in the pathophysiology of pre-eclampsia and related podocyte injury.
    Authors: Craici, Iasmina, Wagner, Steven J, Weissgerber, Tracey L, Grande, Joseph P, Garovic, Vesna D
    Kidney Int, 2014;86(2):275-85.
    Species: Rat
    Sample Types:
  16. A2B adenosine receptors prevent insulin resistance by inhibiting adipose tissue inflammation via maintaining alternative macrophage activation.
    Authors: Csoka B, Koscso B, Toro G, Kokai E, Virag L, Nemeth Z, Pacher P, Bai P, Hasko G
    Diabetes, 2014;63(3):850-66.
    Species: Mouse
    Sample Types: Plasma
  17. FcgammaRIIb inhibits immune complex-induced VEGF-A production and intranodal lymphangiogenesis.
    Authors: Clatworthy M, Harford S, Mathews R, Smith K
    Proc Natl Acad Sci U S A, 2014;111(50):17971-6.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  18. Role of TGF-beta signaling in generation of CD39+CD73+ myeloid cells in tumors.
    Authors: Ryzhov S, Pickup M, Chytil A, Gorska A, Zhang Q, Owens P, Feoktistov I, Moses H, Novitskiy S
    J Immunol, 2014;193(6):3155-64.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  19. The role of IL-1beta in the early tumor cell-induced angiogenic response.
    Authors: Carmi Y, Dotan S, Rider P, Kaplanov I, White M, Baron R, Abutbul S, Huszar M, Dinarello C, Apte R, Voronov E
    J Immunol, 2013;190(7):3500-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  20. Von Hippel-Lindau protein in the RPE is essential for normal ocular growth and vascular development.
    Authors: Lange CA, Luhmann UF, Mowat FM
    Development, 2012;139(13):2340-50.
    Species: Mouse
    Sample Types: Tissue Homogenates
  21. Long-term continuous corticosterone treatment decreases VEGF receptor-2 expression in frontal cortex.
    Authors: Howell KR, Kutiyanawalla A, Pillai A
    PLoS ONE, 2011;6(5):e20198.
    Species: Mouse
    Sample Types: Serum
  22. Tumour hypoxia promotes tolerance and angiogenesis via CCL28 and T(reg) cells.
    Authors: Facciabene A, Peng X, Hagemann IS, Balint K, Barchetti A, Wang LP, Gimotty PA, Gilks CB, Lal P, Zhang L, Coukos G
    Nature, 2011;475(7355):226-30.
    Species: Mouse
    Sample Types: Ascites Fluid
  23. CD11c(hi) dendritic cells regulate the re-establishment of vascular quiescence and stabilization after immune stimulation of lymph nodes.
    Authors: Tzeng TC, Chyou S, Tian S, Webster B, Carpenter AC, Guaiquil VH, Lu TT
    J. Immunol., 2010;184(8):4247-57.
    Species: Mouse
    Sample Types: Tissue Homogenates
  24. Matrix metalloproteinase-9 deficiency worsens lung injury in a model of bronchopulmonary dysplasia.
    Authors: Lukkarinen H, Hogmalm A, Lappalainen U, Bry K
    Am. J. Respir. Cell Mol. Biol., 2009;41(1):59-68.
    Species: Mouse
    Sample Types: Tissue Homogenates
  25. IL-6 loss causes ventricular dysfunction, fibrosis, reduced capillary density, and dramatically alters the cell populations of the developing and adult heart.
    Authors: Banerjee I, Fuseler JW, Intwala AR, Baudino TA
    Am. J. Physiol. Heart Circ. Physiol., 2009;296(5):H1694-704.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  26. Adenosine receptors in regulation of dendritic cell differentiation and function.
    Authors: Novitskiy SV, Ryzhov S, Zaynagetdinov R, Goldstein AE, Huang Y, Tikhomirov OY, Blackburn MR, Biaggioni I, Carbone DP, Feoktistov I, Dikov MM
    Blood, 2008;112(5):1822-31.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  27. CCL3-CCR5 axis regulates intratumoral accumulation of leukocytes and fibroblasts and promotes angiogenesis in murine lung metastasis process.
    Authors: Wu Y, Li YY, Matsushima K, Baba T, Mukaida N
    J. Immunol., 2008;181(9):6384-93.
    Species: Mouse
    Sample Types: Tissue Homogenates
  28. Fibroblast-type reticular stromal cells regulate the lymph node vasculature.
    Authors: Chyou S, Ekland EH, Carpenter AC, Tzeng TC, Tian S, Michaud M, Madri JA, Lu TT
    J. Immunol., 2008;181(6):3887-96.
    Species: Mouse
    Sample Types: Tissue Homogenates
  29. In the adult mesenchymal stem cell population, source gender is a biologically relevant aspect of protective power.
    Authors: Crisostomo PR, Markel TA, Wang M, Lahm T, Lillemoe KD, Meldrum DR
    Surgery, 2007;142(2):215-21.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  30. A functional comparison of canine and murine bone marrow derived cultured mast cells.
    Authors: Lin TY, London CA,
    Vet. Immunol. Immunopathol., 2006;114(3):320-34.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  31. Impaired inflammatory angiogenesis, but not leukocyte influx, in mice lacking TNFR1.
    Authors: Barcelos LS, Talvani A, Teixeira AS, Vieira LQ, Cassali GD, Andrade SP, Teixeira MM
    J. Leukoc. Biol., 2005;78(2):352-8.
    Species: Mouse
    Sample Types: Complex Sample Type
  32. Short-term BMP-2 expression is sufficient for in vivo osteochondral differentiation of mesenchymal stem cells.
    Authors: Noel D, Gazit D, Bouquet C, Apparailly F, Bony C, Plence P, Millet V, Turgeman G, Perricaudet M, Sany J, Jorgensen C
    Stem Cells, 2004;22(1):74-85.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  33. ACE2 activation protects against cognitive decline and reduces amyloid pathology in the Tg2576 mouse model of Alzheimer's disease.
    Authors: Evans C, Miners J, Piva G, Willis C, Heard D, Kidd E, Good M, Kehoe P
    Acta Neuropathol, 0;139(3):485-502.
    Species: Mouse
    Sample Types: Cell Lysates

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Reviews for Mouse VEGF DuoSet ELISA

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Mouse VEGF DuoSet ELISA
By Anonymous on 03/29/2018
Application: Sample Tested: mouse cell conditioned media

Worked perfectly for standards, my samples and a control of diluted recombinant VEGF. Excellent elisa and easy to use


Mouse VEGF DuoSet ELISA
By Anonymous on 05/02/2017
Application: Sample Tested: Cell culture supernatant