Rat VEGF DuoSet ELISA

Catalog # Availability Size / Price Qty
DY564
Ancillary Products Available
Rat VEGF ELISA Standard Curve
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Product Details
Procedure
Citations (11)
FAQs
Supplemental Products
Reviews (1)

Rat VEGF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For 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 rat 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

Rat 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 formation of anti-parallel disulfide-linked dimers (4). Alternately spliced isoforms of 120, 164, and 188 amino acids (aa) have been found in rats and 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 rats and mice. Isoforms other than VEGF120 and VEGF121 contain basic heparin-binding regions and are not freely diffusible (4). Rat VEGF164 shares 97% aa sequence identity with corresponding regions of mouse, 88% with human and bovine, 89% with porcine and canine, and 90% with feline and equine 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). 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, 16, 17). 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 (18). 
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 (19-21). 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 (22, 23). Various strategies have been employed therapeutically to antagonize VEGF-mediated tumor angiogenesis (24). Circulating VEGF levels correlate with disease activity in autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and systemic lupus erythematosus (25).

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 Rat 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.

11 Citations: Showing 1 - 10
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  1. Effects of lncRNA BANCR on endometriosis through ERK/MAPK pathway
    Authors: MB Zhu, LP Chen, M Hu, Z Shi, YN Liu
    Eur Rev Med Pharmacol Sci, 2019;23(16):6806-6812.
    Species: Rat
    Sample Types: Serum
  2. Improvement of diabetic wound healing by topical application of Vicenin-2 hydrocolloid film on Sprague Dawley rats
    Authors: WS Tan, P Arulselvan, SF Ng, CN Mat Taib, MN Sarian, S Fakurazi
    BMC Complement Altern Med, 2019;19(1):20.
    Species: Rat
    Sample Types: Tissue Homogenates
  3. VEGF/Flk1 Mechanism is Involved in Roxarsone Promotion of Rat Endothelial Cell Growth and B16F10 Xenograft Tumor Angiogenesis
    Authors: S Chen, J Xu, Q Wei, Z Zhao, X Chen, H Cui, Y Zhang
    Sci Rep, 2019;9(1):17417.
    Species: Rat
    Sample Types: Cell Culture Supernates
  4. Nrf2 Activation Is a Potential Therapeutic Approach to Attenuate Diabetic Retinopathy
    Authors: D Deliyanti, SF Alrashdi, SM Tan, C Meyer, KW Ward, JB de Haan, JL Wilkinson-
    Invest. Ophthalmol. Vis. Sci., 2018;59(2):815-825.
    Species: Rat
    Sample Types: Cell Lysates
  5. Lifelong environmental enrichment in the absence of exercise protects the brain from age-related cognitive decline
    Authors: AM Birch, ÁM Kelly
    Neuropharmacology, 2018;0(0):.
    Species: Rat
    Sample Types: Tissue Culture Supernates
  6. Albumin suppresses vascular endothelial growth factor via alteration of hypoxia-inducible factor/hypoxia-responsive element pathway.
    Authors: Katavetin P, Inagi R, Miyata T, Tanaka T, Sassa R, Ingelfinger JR, Fujita T, Nangaku M
    Biochem. Biophys. Res. Commun., 2007;367(2):305-10.
    Species: Rat
    Sample Types: Cell Culture Supernates
  7. Transforming growth factor-beta1 enhanced vascular endothelial growth factor synthesis in mesenchymal stem cells.
    Authors: Wang XJ, Dong Z, Zhong XH, Shi RZ, Huang SH, Lou Y, Li QP
    Biochem. Biophys. Res. Commun., 2007;365(3):548-54.
    Species: Rat
    Sample Types: Cell Culture Supernates
  8. Bone marrow molecular alterations after myocardial infarction: Impact on endothelial progenitor cells.
    Authors: Thum T, Fraccarollo D, Galuppo P, Tsikas D, Frantz S, Ertl G, Bauersachs J
    Cardiovasc. Res., 2006;70(1):50-60.
    Species: Rat
    Sample Types: Cell Lysates
  9. Inhibition of angiogenesis by interleukin-4 gene therapy in rat adjuvant-induced arthritis.
    Authors: Haas CS, Amin MA, Allen BB, Ruth JH, Haines GK, Woods JM, Koch AE
    Arthritis Rheum., 2006;54(8):2402-14.
    Species: Rat
    Sample Types: Tissue Homogenates
  10. Prophylactic adenovirus-mediated human kallistatin gene therapy suppresses rat arthritis by inhibiting angiogenesis and inflammation.
    Authors: Wang CR, Chen SY, Wu CL, Liu MF, Jin YT, Chao L, Chao J
    Arthritis Rheum., 2005;52(4):1319-24.
    Species: Rat
    Sample Types: Tissue Homogenates
  11. Effect of tumor host microenvironment on photodynamic therapy in a rat prostate tumor model.
    Authors: Chen B, Pogue BW, Zhou X, O'Hara JA, Solban N, Demidenko E, Hoopes PJ, Hasan T
    Clin. Cancer Res., 2005;11(2):720-7.
    Species: Rat
    Sample Types: Cell Culture Supernates

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Rat VEGF DuoSet ELISA
By Anonymous on 03/25/2020
Application: Sample Tested: Human cell conditioned medium