RayBio® Human VEGF-A ELISA Kit for cell culture supernatants, plasma, serum, and lysate samples.

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Catalog #:
ELH-VEGF
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Product Description

Specifications

Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Species Human
Accession Number P15692
Gene Id 7422
Gene Symbols VEGFA|VEGF
Protein Name / Synonyms Vascular endothelial growth factor A (VEGF-A) (Vascular permeability factor) (VPF)
Quantitative/Semi-Quantitative Quantitative
Specificity This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO.
Compatible Sample Types Cell Culture Supernatants, Plasma, Serum, Tissue Lysates, Cell Lysates
Solid Support 96-well Microplate
Method Of Detection Colorimetric
Design Principle Sandwich-based
Research Area Angiogenesis, Antibody Drug Development, Protein Therapeutics, Biomarker Detection
Sensitivity 10 pg/ml

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Detection Range 10 pg/ml - 6000 pg/ml
Estimated Lead Time 1-2 business days
Shipping Type Blue ice
Storage ≤-20°C

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Target Background

VEGF is a member of the PDGF/VEGF growth factor family that is often found as a disulfide linked homodimer. It is a glycosylated mitogen that specifically acts on endothelial cells and has various effects, including mediating increased vascular permeability, inducing angiogenesis, vasculogenesis and endothelial cell growth, promoting cell migration, and inhibiting apoptosis. Elevated levels of this protein is linked to POEMS syndrome, also known as Crow-Fukase syndrome. Mutations in VEGF gene have been associated with proliferative and nonproliferative diabetic retinopathy.

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Standard Curves

Human VEGF ELISA Kit Standard Curves

Human VEGF ELISA Dilution Series Diagram

Spiking & Recovery Results

VEGF ELISA Spiking and Recovery Results

Linearity Results

VEGF ELISA Linearity Results

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Storage/Stability

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Mause S., Ritzel E., Liehn E., et al. Platelet Microparticles Enhance the Vasoregenerative Potential of Angiogenic Early Outgrowth Cells After Vascular Injury. Circulation . 2010 Aug 3;122(5):495-506. doi: 10.1161/CIRCULATIONAHA.109.909473.
Species:
Human
Sample Type:
Conditioned Media (HUVEC cells in vascular injury model)
De Barros, Sandra, et al. "Aging-related decrease of human ASC angiogenic potential is reversed by hypoxia preconditioning through ROS production." Molecular Therapy 21.2 (2013): 399-408.
Species:
Human
Sample Type:
Conditioned Media (hASCs under normoxic and hypoxic conditions)
Fermoselle C., Rabinovich R., Ausin P., et al. Does oxidative stress modulate limb muscle atrophy in severe COPD patients? Eur Respir J. 2012 Oct;40(4):851-62.
Species:
Human
Sample Type:
Tissue Lysate (Quadricep Biopsies from COPD patients)
Bigalke C., et al. VEGF-releasing suture material for enhancement of vascularization: 4 Development, in vitro and in vivo study. Acta Biomaterialia 7 September 2014, DOI: 10.1016/j.actbio.2014.09.002
Species:
Human
Sample Type:
Serum
Diaz-Gomez L., Concheiro A., Alvarez-Lorenzo C., Garcia-Gonzalez C. Growth factors delivery from hybrid PCL-starch scaffolds processed using supercritical fluid technology. Carbohydr Polym. 2016 May 20;142:282-92. doi: 10.1016/j.carbpol.2016.01.051.
Species:
Human
Sample Type:
Conditioned Media (PCL-Starch scaffolds)
Paget, M. B., et al. "Rotational co?culture of clonal ??cells with endothelial cells: effect of PPAR?? agonism in vitro on insulin and VEGF secretion." Diabetes, Obesity and Metabolism 13.7 (2011): 662-668.
Species:
Human
Sample Type:
Conditioned Media
Wang XS, Williams LA, Krishnan S, et al. Serum sTNFR1, IL6, and the Development of Fatigue in Patients with Gastrointestinal Cancer Undergoing Chemoradiation Therapy. Brain, Behavior, and Immunity. 2012;26(5):699-705. doi:10.1016/j.bbi.2011.12.007.
Species:
Human
Sample Type:
Serum (Colorectal or esphageal cancer patients treated with concurrent chemoradiation therapy)
Wang, Y., Lan, H., Yin, T., Zhang, X., Huang, J., Fu, H., Wang, Z. (2019). Covalent immobilization of biomolecules on stent materials through mussel adhesive protein coating to form biofunctional films. Materials Science and Engineering: C, 110187. doi:10.1016/j.msec.2019.110187 
Species:
Human
Sample Type:
Conditioned media (VEGF in mixing solutions)
Platt, DH et al. Peroxynitrite increases VEGF expression in vascular endothelial cells via STAT3. Peroxynitrite increases VEGF expression in vascular endothelial cells via STAT3. Peroxynitrite increases VEGF expression in vascular endothelial cells via STAT3. Free Radic. Biol. Med. Volume 39, Issue 10, Pages 1353-61, Nov 2005
Species:
Bovine
Sample Type:
Cell Lysate (Cell cultures treated with peroxynitrite)
Etulain, J., et al. "Hyperthermia inhibits platelet hemostatic functions and selectively regulates the release of alpha?granule proteins." Journal of Thrombosis and Haemostasis 9.8 (2011): 1562-1571.
Species:
Human
Sample Type:
Conditioned Media (platelets)
  1. Easy to use
    Rating
    100%
    We have been using this kit for many years. Very easy to use.

    Review by

    from Medical University of SC,

    on

  2. Great product
    Rating
    100%
    Very easy to use. Excellent results.

    Review by

    from Michigan State University,

    on

  3. Great product
    Rating
    100%
    It is a very user-friendly product for people who work with this type of sample for the first time, highly recommended.

    Review by

    from University of Puerto Rico,

    on

  4. Great product
    Rating
    100%
    This kit is simple to use and ideal for those who are new to evaluating factors on culture media. All directions are simple to follow, and the reagents are high quality.

    Review by

    from University of Puerto Rico at Mayaguez,

    on

  5. Great product
    Rating
    100%
    Very good sensitivity

    Review by

    from BCM,

    on

  6. Great product
    Rating
    100%
    Had great success using these kits. Really clear results and effective. These kits are easy to use and come with all the components needed which is a plus! Look forward to using them again.

    Review by

    from Advirna,

    on

  7. Great product
    Rating
    100%
    I've used this product several times and am very happy with its high sensitivity, ease of use, and the technical support.

    Review by

    from State University of New York,

    on

  8. Great product
    Rating
    80%
    I used cell culture supernatant and it was really easy to perform

    Review by

    from CPMC,

    on

  9. Great product
    Rating
    100%
    Use it to test in vitro releasing of VEGF-A from tissue engineering scaffold. Easy to use and work very well.

    Review by

    from North Carolina State University,

    on

  10. Great product
    Rating
    80%
    I used this kit for first time and works really good

    Review by

    from CPMC,

    on

  11. Great product
    Rating
    100%
    Very sensitive kit, instructions are easy to follow and have yielded great results!

    Review by

    from University of Illinois,

    on

  12. Great product
    Rating
    100%
    Writing another review because we have had nothing but success with this kit! Looking to publish the results soon!

    Review by

    from University of Florida,

    on

  13. Great product
    Rating
    100%
    Easy to use!

    Review by

    from University of San Diego,

    on

  14. Great product
    Rating
    100%
    The ELISA instructions are concise and easy to follow. The kit also has all reagents you need. It has great reproducibility and reliability.

    Review by

    from University of Florida,

    on

  15. Great product
    Rating
    100%
    This product has been reliable with a descriptive protocol booklet that is easy to follow.

    Review by

    from Immutrix Therapeutics,

    on

  16. Great product
    Rating
    100%
    Very simple, concise instructions. This ELISA kit has a wide concentration detection range. It performed as intended and will continue using their ELISA kits. I fully recommend RayBiotech to anyone in lab.

    Review by

    from Stanford University,

    on

  17. Great product
    Rating
    100%
    I'm really impressed by all Raybiotech ELISA kits. This one is not an exception: it works well with tumor tissue lysates, and allows to analyze tumor cell-derived VEGF-A in xenograft mouse models.

    Review by

    from Roswell Park Comprehensive Cancer Center,

    on

  18. Great product
    Rating
    80%
    We ordered 2 boxes of this kit (along side with 2 of Periostin and 2 of IL-4 ELISA kits). The kits came in strong, sturdy boxes. The instructions are clear and easy to follow. The only thing is we wish that the sensitivity was lower. The current minimum detectable dose of Human VEGFA is 10 pg/ml. We had to dilute the standards and remove one of the them from the standard curve to be able to detect the protein levels. But we used these kits on urine samples and it seemed like VEGFA was very low in urine anyways. Just an idea for future improvement.

    Review by

    from New York Medical College,

    on

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